WO2014184655A1

WO2014184655A1 - Methods for using crustacean phospholipid-peptide-protein complexes

Info

Publication number: WO2014184655A1
Application number: PCT/IB2014/001068
Authority: WO
Inventors: Inge Bruheim; Mikko Griinari; Stig Rune REMOY; Even REMOY
Original assignee: Olympic Seafood AS
Current assignee: Olympic Seafood AS
Priority date: 2013-02-07
Filing date: 2014-02-06
Publication date: 2014-11-20
Anticipated expiration: 2015-08-07

Abstract

The present invention relates to treatments of medical disorders with crustacean phospholipid-peptide-protein (PPC) compositions. For example, such a composition can be a low fluoride composition, a low trimethylamine composition or a composition that is low in both fluoride and trimethylamine. The PPC compositions may be further supplemented with additional ingredients including, but not limited to, low fluoride crustacean oil (e.g., low fluoride krill oil), omega-3 fatty acids, vitamins, minerals and/or antioxidants.

Description

Methods For Using Crustacean Phospholipid-Peptide-Protein Complexes

Field of the Invention

The present invention is related to high quality crustacean phospholipid-peptide- protein (PPC) compositions for use as nutritional supplements, cosmetic products, pharmaceuticals and clinical nutrition. For example, the use of crustacean PPC compositions can treat medical disorders and/or to improve mental and/or physical performance. In particular, PPC may comprise a phospholipid-peptide-protein complex or a de-oiled phospholipid-peptide-protein complex. Clinically, such crustacean PPC compositions can improve symptoms of various medical disorders.

Background of the Invention

Krill are marine crustaceans (Class Malacostraca, Order Euphausiacea) comprising approximately 86 species, a majority of which are free swimming, and are considered plankton (Everson 2000). Krill sometimes form dense swarms that can extend over several square kilometers and represent a biomass of thousands or even millions of tons (Nicol and Endo 1999).

There are currently several active krill fisheries but these are dominated by two; one based in Antarctica for E. superba and the other based predominantly in Japan (but also Canada) targeting E. pacifica. Together these two fisheries E. superba and E. pacified) represent at least 97% of the total krill landed. The low levels of environmental pollutants in the Antarctic krill is a benefit for the utilization of the krill for health products. Currently available krill products for human consumption is mainly based on krill oil in where the protein fraction is removed.

To utilize the whole krill for nutritional supplements or for pharmaceuticals (clinical nutrition) there is a need for compositions and formulations in where most of the nutrients and the bioactive components from krill are kept intact and where both lipid soluble and lipid insoluble micronutrients which are required can be in mixed in a feasible way.

The fast growing elderly population and expectation of longer lifespan increase the importance of keeping elderly population healthy. Due to thi s, there is a great need for effective nutritional supplements that the eldery can easily digest and absorb that contains all essential nutrients, in addition to micronutrients and health ingredients. In particular, what is needed is a nutraceutical composition able to reduce low grade chronic inflammation and g oxidative stress. This would be beneficial for the elderly population and for prevention of age related diseases.

Summary Of The Invention

The present invention is related to high quality crustacean phospholipid-peptide- protein (PPC) compositions for use as nutritional supplements, cosmetic products, pharmaceuticals and clinical nutrition. For example, the use of cmstacean PPC compositions can treat medical disorders and/or to improve mental and/or physical performance. In particular, PPC may comprise a phospholipid-peptide-protein complex or a de-oiled phospholipid-peptide-protein complex. Clinically, such crustacean PPC compositions can improve symptoms of various medical disorders.

In one embodiment, the present invention contemplates a method, comprising: a) providing: i) a patient exhibiting at least one symptom of a medical disorder; ii) a low fluoride crustacean meal formulation; b) administering said composition to said patient under conditions such that said at least one symptom is reduced. In one embodiment the medical disorder comprises an age-related medical disorder. In one embodiment, the age-related medical disorder includes, but is not limited to, a lack of homeostatic control, macular degeneration, diabetes, or inflammation. In one embodiment, the medical disorder comprises malnutrition, hi one embodiment, the medical disorder comprises an ocular disorder. In one embodiment, the medical disorder comprises a cardiovascular disorder. In one embodiment, the medical disorder comprises a skeletal medical disorder. In one embodiment, the medical disorder comprises a central nervous system disorder. In one embodiment, the medical disorder comprises a muscular disorder. In one embodiment the medical disorder comprises cachexia. In one embodiment, the medical disorder comprises digestive tract medical disorder. In one embodiment, the medical disorder comprises a dyslipidemic medical disorder. In one embodiment, the medical disorder comprises a hair disorder. In one embodiment, the medical disorder comprises a nail disorder. In one embodiment, the medical disorder comprises a skin disorder. In one embodiment, the medical disorder comprises a sexual disorder. In one embodiment, the formulation is a pharmaceutically acceptable formulation. In one embodiment, the crustacean meal formulation is a krill meal formulation. In one embodiment the crustacean meal formulation is a medicament. In one embodiment, the meal formulation comprises a low fluoride phospholipid-peptide-protein complex (e.g., for example, Krill Meal 1). In one embodiment, the meal formulation comprises a low fluoride de-oiled phospholipid-protein complex (e.g., for example, Krill Meal 2). In one embodiment, the meal formulation further comprises a low fluoride krill oil. In one embodiment, the meal formulation further comprises a low trimethyl amine krill oil. In one embodiment, the meal formulation further comprises a low fluoride, low trimethyl amine krill oil. hi one embodiment, the meal formulation further comprises a low fluoride extraction residue. In one embodiment, the meal formulation further comprises an additional ingredient including, but not limited to, minerals, lipid soluble vitamins, lipid insoluble vitamins, bioactive health ingredients and/or omega-3 oils. In one embodiment, the administered composition ranges between 0.005 - 0.50 grams per day per kilogram of said patient's body weight, hi one embodiment, the formulation includes, but is not limited to, a capsule, a gelatin capsule, a flavored capsule, a tablet, coated tablets, powders, granulates, solutions, dispersions, suspensions, syrups, emulsions, a liquid nutrition composition and/or a beverage. In one embodiment, the formulation includes, but is not limited to a nutritional supplement, a pharmaceutical, a food supplement, a functional food ingredient, a food additive and/or a nutritional supplement preparation.

In one embodiment, the present invention contemplates a method, comprising: a) providing; i) a patient exhibiting at least one symptom of an ocular medical disorder; ii) a crustacean meal formulation comprising a low fluoride phospholipid-peptide-protein complex, zinc oxide, vitamin C and vitamin E; and b) administering said formulation to said patient under conditions such that said at least one symptom is reduced. In one embodiment, the ocular medical disorder is a retinal disorder. In one embodiment, the ocular medical disorder is macular degeneration. In one embodiment, the formulation is a pharmaceutically acceptable formulation. In one embodiment, the crustacean meal formulation is a krill meal formulation. In one embodiment, the crustacean meal formulation further comprises a low fluoride carstacean oil. In one embodiment, the crustacean meal formulation further comprises a low trimethyl amine crustacean oil. In one embodiment, the meal formulation further comprises a low fluoride extraction residue. In one embodiment, the at least one additional ingredient is selected from at least one of the group comprising minerals, lipid soluble vitamins, lipid insoluble vitamins, bioactive health ingredients and/or omega-3 oils. In one embodiment, the administering comprises a daily effective dose ranging between approximately 1 -3 grams.

In one embodiment, the present invention contemplates a method, comprising: a) providing; i) a patient exhibiting at least one symptom of a cardiovascular medical disorder; ii) a crustacean meal formulation comprising a low fluoride phospholipid-peptide-protein complex, phytosterols, phytostarrnols and vitamin D; and b) administering said formulation to said patient under conditions such that said at least one symptom is reduced. In one embodiment, the cardiovascular medical disorder is a heart disorder. In one embodiment, the formulation is a pharmaceutically acceptable formulation. In one embodiment, the crustacean meal formulation is a krill meal formulation. In one embodiment, the crustacean meal formulation further comprises a low fluoride crustacean oil. In one embodiment, the crustacean meal formulation further comprises a low trimethyl amine crustacean oil. In one embodiment, the meal formulation further comprises a low fluoride extraction residue. In one embodiment, the at least one additional ingredient is selected from at least one of the group comprising minerals, lipid soluble vitamins, lipid insoluble vitamins, bioactive health ingredients and/or omega-3 oils. In one embodiment, the administering comprises a daily effective dose ranging between approximately 0.5 -4 grams.

In one embodiment, the present invention contemplates a method, comprising: a) providing; i) a patient exhibiting at least one symptom of a skeletal medical disorder; ii) a crustacean meal formulation comprising a low fluoride phospholipid-peptide-protein complex, calcium carbonate, vitamin D and vitamin K; and b) administering said formulation to said patient under conditions such that said at least one symptom is reduced. In one embodiment, the skeletal medical disorder is oestoporosis. In one embodiment, the formulation is a pharmaceutically acceptable formulation. In one embodiment, the crustacean meal formulation is a krill meal formulation. In one embodiment, the crustacean meal formulation further comprises a low fluoride crustacean oil. In one embodiment, the crustacean meal formulation further comprises a low trmethyl amine crustacean oil. In one embodiment, the meal formulation further comprises a low fluoride extraction residue. In one embodiment, the at least one additional ingredient is selected from at least one of the group comprising minerals, lipid soluble vitamins, lipid insoluble vitamins, bioactive health ingredients and/or omega-3 oils. In one embodiment, the administering comprises a daily effective dose ranging between approximately 0.5 - 4 grams.

In one embodiment, the present invention contemplates a method, comprising: a) providing; i) a patient exhibiting at least one symptom of a central nervous system medical disorder; ii) a crustacean meal formulation comprising a low fluoride phospholipid-peptide- protein complex and ubiqinon (e.g., for example, Q10); and b) administering said formulation to said patient under conditions such that said at least one symptom is reduced. In one embodiment, the central nervous system medical disorder is dementia. In one embodiment, the central nervous system medical disorder is multiple sclerosis. In one embodiment, the central nervous system medical disorder is reduced mental acuity. In one embodiment, the centeral nervous system medical disorder is reduced concentration. In one embodiment, the formulation is a pharmaceutically acceptable formulation. In one embodiment, the crustacean meal formulation is a krill meal formulation. In one embodiment, the crustacean meal formulation further comprises a low fluoride crustacean oil. In one embodiment, the crustacean meal formulation further comprises a low trimethyl amine crustacean oil. In one embodiment, the meal formulation further comprises a low fluoride extraction residue. In one embodiment, the at least one additional ingredient is selected from at least one of the group comprising minerals, lipid soluble vitamins, lipid insoluble vitamins, bioactive health ingredients and/or omega-3 oils. In one embodiment, the administering comprises a daily effective dose ranging between approximately 1 -3 grams.

In one embodiment, the present invention contemplates a method, comprising: a) providing; i) a patient exhibiting at least one symptom of an ocular medical disorder; ii) a crustacean meal formulation comprising a low fluoride de-oiled phospholipid-peptide complex, zinc oxide, vitamin C, vitamin E, lutein, zeaxanthin and DHA; and b) administering said formulation to said patient under conditions such that said at least one symptom is reduced. In one embodiment, the ocular medical disorder is a retinal disorder. In one embodiment, the ocular medical disorder is macular degeneration. In one embodiment, the formulation is a pharmaceutically acceptable formulation. In one embodiment, the crustacean meal formulation is a krill meal formulation. In one embodiment, the crustacean meal formulation further comprises a low fluoride crustacean oil. In one embodiment, the crustacean meal formulation further comprises a low trimethyl amine crustacean oil. In one embodiment, the meal formulation further comprises a low fluoride extraction residue. In one embodiment, the at least one additional ingredient is selected from at least one of the group comprising minerals, lipid soluble vitamins, lipid insoluble vitamins, bioactive health ingredients and/or omega-3 oils. In one embodiment, the administering comprises a daily effective dose ranging between approximately 1 -3 grams.

In one embodiment, the present invention contemplates a method, comprising: a) providing; i) a patient exhibiting at least one symptom of a muscular medical disorder; ii) a crustacean meal formulation comprising a low fluoride de-oiled phospholipid- protein complex and omega-3 fatty acids; and b) administering said formulation to said patient under conditions such that said at least one symptom is reduced. In one embodiment, the muscular medical disorder is an anabolic medical disorder. In one embodiment, the anabolic medical disorder is a mitochondrial disorder. In one embodiment, the formulation is a

pharmaceutically acceptable formulation. In one embodiment, the crustacean meal formulation is a krill meal formulation. In one embodiment, the crustacean meal formulation further comprises a IOAV fluoride crustacean oil. In one embodiment, the crustacean meal formulation further comprises a low trimethyl amine crustacean oil. In one embodiment, the meal formulation further comprises a low fluoride extraction residue. In one embodiment, the at least one additional ingredient is selected from at least one of the group comprising minerals, lipid soluble vitamins, lipid insoluble vitamins, bioactive health ingredients and/or omega-3 oils. In one embodiment, the admimstering comprises a daily effective dose ranging between approximately 1 -3 grams.

In one embodiment, the present invention contemplates a method, comprising: a) providing; i) a patient exhibiting at least one symptom of a cachexic medical disorder; ii) a crustacean meal formulation selected from the group consisting of a low fluoride

phospholipid-peptide-protein complex and a low fluoride de-oiled phospholipid-protein complex and at least one additional ingredient; and b) administering said formulation to said patient under conditions such that said at least one symptom is reduced. In one embodiment, the cachexic medical disorder is cancer. In one embodiment, the cachexic medical disorder is malnutrition. In one embodiment, the cachexic medical disorder is a chronic infectious disease. In one embodiment, the formulation is a pharmaceutically acceptable formulation, hi one embodiment, the crustacean meal formulation is a krill meal formulation. In one embodiment, the crustacean meal formulation further comprises a low fluoride crustacean oil. In one embodiment, the crustacean meal formulation further comprises a low trimethyl amine crustacean oil. In one embodiment, the meal formulation further comprises a low fluoride extraction residue. In one embodiment, the at least one additional ingredient is selected from at least one of the group comprising minerals, lipid soluble

lipid insoluble vitamins, bioactive health ingredients and/or omega-3 oils. In one embodiment, the administering comprises a daily effective dose ranging between approximately 0.5 -4 grams.

In one embodiment, the present invention contemplates a method, comprising: a) providing; i) a patient exhibiting at least one symptom of a lack of homestatic control; ii) a crustacean meal formulation selected from the group consisting of a low fluoride

phospholipid-peptide-protein complex and a low fluoride de-oiled phospholipid-protein complex and at least one additional ingredient; and b) admimstering said formulation to said patient under conditions such that said at least one symptom is reduced. In one embodiment, the formulation is a pharmaceutically acceptable formulation, i one embodiment, the crustacean meal formulation is a krill meal formulation. In one embodiment, the crustacean meal formulation further comprises a low fluoride crustacean oil. In one embodiment, the crustacean meal formulation further comprises a low trimethyl amine crustacean oil. In one embodiment, the meal formulation further comprises a low fluoride extraction residue. In one embodiment, the at least one additional ingredient is selected from at least one of the group comprising minerals, lipid soluble vitamins, lipid insoluble vitamins, bioactive health ingredients and/or omega-3 oils. In one embodiment, the adm stering comprises a daily effective dose ranging between approximately 0.5 -4 grams.

In one embodiment, the present invention contemplates a method, comprising: a) providing; i) a patient exhibiting at least one symptom of an endocrinologic medical disorder; ii) a crustacean meal formulation selected from the group consisting of a low fluoride phospholipid-pep tide-protein complex and a low fluoride de-oiled phospholipid-protein complex and at least one additional ingredient; and b) administering said formulation to said patient under conditions such that said at least one symptom is reduced. In one embodiment, the endocrinologic medical disorder is diabetes. In one embodiment, the formulation is a pharmaceutically acceptable formulation. In one embodiment, the crustacean meal formulation is a krill meal formulation. In one embodiment, the crustacean meal formulation further comprises a low fluoride crustacean oil. In one embodiment, the crustacean meal formulation further comprises a low trimethyl amine crustacean oil. In one embodiment, the meal formulation further comprises a low fluoride extraction residue, hi one embodiment, the at least one additional ingredient is selected from at least one of the group comprising minerals, lipid soluble vitamins, lipid insoluble vitamins, bioactive health ingredients and/or omega-3 oils. In one embodiment, the administering comprises a daily effective dose ranging between approximately 0.5 -4 grams.

In one embodiment, the present invention contemplates a method, comprising: a) providing; i) a patient exhibiting at least one symptom of an inflammatory medical disorder; ii) a crustacean meal formulation selected from the group consisting of a low fluoride phospholipid-peptide-protein complex and a low fluoride de-oiled phospholipid-protein complex and at least one additional ingredient; and b) administering said formulation to said patient under conditions such that said at least one symptom is reduced. In one embodiment, the inflammatory medical disorder is arthritis. In one embodiment, the formulation is a pharmaceutically acceptable formulation. In one embodiment, the crustacean meal formulation is a krill meal formulation. In one embodiment, the crustacean meal formulation further comprises a low fluoride crustacean oil. In one embodiment, the crustacean meal formulation further comprises a low trimethyl amine crustacean oil. In one embodiment, the meal formulation further comprises a low fluoride extraction residue. In one embodiment, the at least one additional ingredient is selected from at least one of the group comprising minerals, lipid soluble vitamins, lipid insoluble vitamins, bioactive health ingredients and/or omega-3 oils. In one embodiment, the administering comprises a daily effective dose ranging between approximately 0.5 -4 grams.

In one embodiment, the present invention contemplates a method, comprising: a) providing; i) a patient exhibiting at least one symptom of a digestive tract medical disorder; ii) a crustacean meal formulation selected from the group consisting of a low fluoride phospholipid-peptide-protein complex and a low fluoride de-oiled phospholipid-protein complex and at least one additional ingredient; and b) administering said formulation to said patient under conditions such that said at least one symptom is reduced. In one embodiment, the digestive tract medical disorder is irritable bowel disease. In one embodiment, the digestive tract medical disorder is Crohn's disease. In one embodiment, the digestive tract medical disorder is ulcerative colitis, i one embodiment, the formulation is a

pharmaceutically acceptable formulation. In one embodiment, the crustacean meal formulation is a krill meal formulation. In one embodiment, the crustacean meal formulation further comprises a low fluoride crustacean oil. In one embodiment, the crustacean meal formulation further comprises a low trimethyl amine crustacean oil. In one embodiment, the meal formulation further comprises a low fluoride extraction residue. In one embodiment, the at least one additional ingredient is selected from at least one of the group comprising minerals, lipid soluble vitamins, lipid insoluble vitamins, bioactive health ingredients and/or omega-3 oils. In one embodiment, the administering comprises a daily effective dose ranging between approximately 0.5 -4 grams.

In one embodiment, the present invention contemplates a method, comprising: a) providing; i) a patient exhibiting at least one symptom of a dyslipidemic medical disorder; ii) a crustacean meal formulation selected from the group consisting of a low fluoride phospholipid-peptide-protein complex and a low fluoride de-oiled phospholipid-protein complex and at least one additional ingredient; and b) administering said formulation to said patient under conditions such that said at least one symptom is reduced. In one embodiment, the dyslipidemic medical disorder is elevated low density lipoprotein cholesterol levels, hi one embodiment, the dyslipidemic medical disorder is reduced high density lipoprotein cholestrol levels, hi one embodiment, the dyslipidemic medical disorder is elevated triglyceride levels. In one embodiment, the formulation is a pharmaceutically acceptable fonnulation. In one embodiment, the crustacean meal formulation is a krill meal formulation. In one embodiment, the crustacean meal formulation further comprises a low fluoride crustacean oil. In one embodiment, the crustacean meal formulation further comprises a low trimethyl amine crustacean oil. In one embodiment, the meal formulation further comprises a low fluoride extraction residue. In one embodiment, the at least one additional ingredient is selected from at least one of the group comprising minerals, lipid soluble vitamins, lipid insoluble vitamins, bioactive health ingredients and/or omega-3 oils. In one embodiment, the administeiing comprises a daily effective dose ranging between approximately 0.5 -4 grams.

In some embodiments of the above methods, the formulation further comprises a lipid insoluble ingredient. In one embodiment, the lipid insoluble ingredient comprises a lipid insoluble vitamin. In one embodiment, the lipid insoluble vitamin comprises approximately between 5 - 50 wt% of the formulation. In one embodiment, the lipid insoluble vitamin includes, but is not limited to, vitamin C, vitamin B12 and/or folate. In one embodiment, the lipid insoluble ingredient comprises cognitive health protecting substances. In one embodiment, the cognitive health protecting substances include, but are not limited to, resveratrol, astaxanthin or ubiquinone. In one embodiment, the lipid insoluble ingredient comprises cartilage protecting substances. In one embodiment, the cartilage protecting substances are glucosamine and chondroitin. In one embodiment, the lipid insoluble ingredients are selected from the group consisting of glucosamine hydrochloride, glucosamine sulfate, glucosamine potassium, glucosamine sodium, N-acetyl d-glucosamine, chondroitin, chondroitin sulfate, curcumin, ascorbic acid, hyaluronic acid, green lipped mussel powder, creatine, L-carnitine, ascorbic acid, manganese, manganese proteinate, zinc, zinc proteinate, copper, copper proteinate, ginseng, green tea extract, ginger, garlic, vincamine, grape seed extract, grape seed meal, dimethyl glycine, whey protein, brewer's yeast, St. John's wort, vinpocetine, aloe vera, ginko biloba, curcumin, betaglucans, and mannaoligosaccharides. In one embodiment, the weight ratio of krill meal to lipid insoluble ingredients in the compo sition ranges between approximately 16:1 to 1 : 1.

In some embodiments of the above methods, the formulation further comprises a lipid soluble vitamin. In one embodiment, the lipid soluble vitamin includes, but is not limited to, vitamin E, vitamin A and/or vitamin D. In one embodiment, the lipid soluble vitamin comprises approximately between 5 - 50 wt% of the formulation. In one embodiment, the lipid soluble ingredient is selected from the group consisting of vitamin A, vitamin D, vitamin E, alpha lipoic acid, lutein, and natural carotenoids. In one embodiment, the composition comprises 10 - 90 % (w/w) krill meal, 10 - 90 % (w/w) lipid insoluble mgredient(s), and 0 - 80 % (w/w) excipients. In some embodiments of the above methods, the formulation further comprises a nutritionally essential mineral. In one embodiment, the nutritionally essential mineral includes, but is not limited to, zinc, calcium, magnesium and/or zinc oxide. In one embodiment, the nutritionally essential mineral comprises approximately between 0.001 - 50 wt% of the formulation.

In some embodiments of the above methods, the formulation further comprises a bioactive health ingredient. In one embodiment, the bioactive health ingredient includes, but is not limited to, zeaxanthin, resveratrol and/or ubiquinon. In one embodiment, the bioactive health ingredient comprises approximately between 0.001 - 50 wt% of the formulation.

In some embodiments of the above methods, the formulation further comprises an omega-3 enriched marine oil. In one embodiment, the omega-3 enriched marme oil includes, but is not limited to, fish oil and/or squid oil. In one embodiment, the omega-3 enriched marine oil is microencapsulated. In one embodiment, the omega-3 enriched marine oil comprises approximately between 5 - 95 wt% of the formulation. In one embodiment, the omega-3 oil is in a form including, but not limited to, an ethylester, a triglyceride and/or a phospholipid.

In some embodiments of the above methods, the formulation further comprises a DHA enriched marine oil. In one embodiment, the DHA enriched marine oil includes, but is not limited to, fish oil and/or squid oil. In one embodiment, the DHA enriched marine oil is microencapsulated. In one embodiment, the DHA enriched marine oil comprises between approximately 5 - 95 wt% of the formulation.

In some embodiments of the above methods, the formulation further comprises an EPA enriched marine oil. i one embodiment, the EPA enriched marine oil includes, but is not limited to, fish oil and/or squid oil. In one embodiment, the EPA enriched marine oil is microencapsulated. In one embodiment, the EPA enriched marine oil comprises bteween approximately 5 - 95 wt% of the formulation.

In some embodiments of the above methods, the formulation further comprises conventional excipients including, but not limited to, solvents, diluents, binders, sweeteners, aromas, pH modifiers, viscosity modifiers, antioxidants, corn starch, lactose, glucose, microcrystalline cellulose, magnesium stearate, tartaric acid, water, ethanol, glycerol, sorbitol, and/or carboxymethyl cellulose. In one embodiment, the excipient is selected from the group consisting of, fillers, granulating agents, adhesives, disintegrants, lubricants, antiadherants, glidants, wetting agents, dissolution retardants, enhancers, adsorbents, buffers, chelating agents, preservatives, colours, flavours, sweeteners, starch, pregelatinized starch, malto dextrin, monohydrous dextrose, alginic acid, sorbitol, mannitol, magnesium stearate, stearic acid, talc, silic, cellulose, microcrystalline cellulose, methyl cellulose,

polyvinylpyrrolidone and - commercial products, such as Aerosil^®, Kollidon^® and Explotab^®.

In some embodiments of the above methods, the administering comprises an effective daily dose of the formulation between approximately 100 mg - 100 gram. In one embodiment, the duration of the administration comprised at least two weeks.

In one embodiment, the present invention contemplates a method, comprising: a) providing: i) an animal exhibiting at least one symptom of a degenerative joint disease; ii) a veterinary supplement comprising a crustacean meal formulation selected from the group consisting of a low fluoride phospholipid-peptide-protein complex and a low fluoride de- oiled phospholipid-protein complex and at least one additional ingredient; b) administering said composition to said animal under conditions such that said at least one symptom is reduced. In one embodiment, the composition is homogenous. In one embodiment, the crustacean meal formulation is a krill meal formulation. In one embodiment, the crustacean meal formulation further comprises a low fluoride crustacean oil. In one embodiment, the crustacean meal formulation further comprises a low trimethyl amine crustacean oil. In one embodiment, the at least one additional ingredient is selected from at least one of the group comprising minerals, lipid soluble vitamins, lipid insoluble vitamins, bioactive health ingredients and/or omega-3 oils. In one embodiment, the at least one symptom comprises pain. In one embodiment, the at least one symptom comprises stiffness. In one embodiment, the at least one symptom comprises lameness. In one embodiment, the at least one symptom comprises mobility. In one embodiment, the at least one symptom comprises mood. In one embodiment, the at least one symptom comprises activity. In one embodiment, the at least one embodiment comprises play. In one embodiment, mobility is assessed by a modified Helsinki Chronic Pain Index. In one embodiment, the lameness is determined by a deviation of a 60:40 weight distribution between the front legs and the rear legs of said animal. In one embodiment the animal is a quadruped animal. In one embodiment, the quadruped animal is selected from at least one of the group comprising a canine, a feline, an equine, a bovine, an ovine, and/or a porcine. In one embodiment, the administering comprises a daily effective dose ranging between approximately 0.005 - 0.50 grams per day per kilogram of said animal's body weight. In one embodiment, the duration of the administration comprised at least six weeks. In one embodiment, the degenerative joint disease comprises osteoarthritis. In one embodiment, the crustacean meal further comprises cartilage protecting substances. In one embodiment, the cartilage protecting substances are glucosamine and chondroitin. In one embodiment, the at least one additional ingredient is selected from the group comprising glucosamine hydrochloride, glucosamine sulfate, glucosamine potassium, glucosamine sodium, N-acetyl d- glucosamine, chondroitin, chondroitin sulfate, curcumin, ascorbic acid, hyaluronic acid, green lipped mussel powder, creatine, L-carnitine, ascorbic acid, manganese, manganese proteinate, zinc, zinc proteinate, copper, copper proteinate, ginseng, green tea extract, ginger, garlic, vincamine, grape seed extract, grape seed meal, dimethyl glycine, whey protein, brewer's yeast, St. John's wort, vinpocetine, aloe vera, ginko biloba, curcumin, betaglucans, and mannaoligosaccharides. In one embodiment, the composition comprises a krill meal to lipid insoluble ingredients weight ratio ranging between approximately 16:1 to 1 : 1. In one embodiment, the composition further comprises a lipid soluble ingredient. In one embodiment, the lipid soluble ingredient is selected from the group consisting of vitamin A, vitamin D, vitamin E, alpha lipoic acid, lutein, and natural carotenoids. In one embodiment, the composition comprises 10 - 90 % (w/w) krill meal, 10 - 90 % (w/w) lipid insoluble ingredient(s), and 0 - 80 % (w/w) excipients. In one embodiment, the composition comprises a suitable form selected from the group consisting of a tablet, a capsule, a granule, a pellet, a powder, or a pet treat. In one embodiment, the capsule comprises a hard gelatin capsule. In one embodiment, the hard gelatin capsule is a sprinkle capsule. In one embodiment, the pet treat comprises at least one flavor ingredient. In one embodiment, the pet treat comprises a filler ingredient. In one embodiment, the composition further comprises an excipient. In one embodiment, the excipient is selected from the group consisting of diluents, fillers, binders, granulating agents, adhesives, disintegrants, lubricants, antiadherants, glidants, wetting agents, dissolution retardants, enhancers, adsorbents, buffers, chelating agents, preservatives, colours, flavours, sweeteners, starch, pregelatinized starch, maltodextrin, monohydrous dextrose, alginic acid, sorbitol, mannitol, magnesium stearate, stearic acid, talc, silic, cellulose, microcrystalline cellulose, methyl cellulose, polyvinylpyrrolidone and - commercial products, such as Aerosil^®, Kollidon^® and Explotab^®.

In one embodiment, the present invention contemplates a method, comprising: a) providing: i) an animal in need of increased food intake; ii) an animal food or feed comprising between approximately 0.01 % (w/w) and 1.0% (w/w) krill meal; b) feeding said animal said food under conditions such that said animal's food intake increases. In one embodiment, the krill meal comprises krill oil. In one embodiment, the krill oil comprises at least one omega-3 fatty acid. In one embodiment, the method further comprises improving the health of said animal. In one embodiment, the animal comprises a non-human animal. In one embodiment, the non-human animal comprises a dog. Definitions

As used herein and in the appended claims, the singular forms "a," "an," and "the" include plural reference unless the context clearly dictates otherwise, e.g., reference to "a method" includes a plurality of such methods.

The words "comprise", "comprises", and "comprising" are to be interpreted inclusively rather than exclusively.

The term "optionally" means here the same as "possibly". For example, compositions disclosed herein as "optionally comprises excipients", means that the composition may or may not comprise excipients, in other words the composition possibly comprises excipients.

The term "patient", as used herein, is a human or animal and need not be hospitalized. For example, out-patients, persons in nursing homes are "patients." A patient may comprise any age of a human or non-human animal and therefore includes both adult and juveniles (i.e., children). It is not intended that the term "patient" connote a need for medical treatment, therefore, a patient may voluntarily or involuntarily be part of experimentation whether clinical or in support of basic science studies.

The term "animal" as used herein, means species including but not limited to mammals, fish, crustaceans, amphibians, reptiles etc. In particular, a "companion animal" refers to any non-human animal kept by a human as a pet or any animal of a variety of species that have been widely domesticated as pets, such as dogs (Canis familiar is), and cats (Felis domesticus), whether or not the animal is kept solely or partly for companionsl ip. Companion animals also include working animals including but not limited to horses, cows, pigs, goats, sheep, dogs (i.e., for example, livestock herding) and/or cats (i.e., for example, rodent control).

The term "effective amount" as used herein, refers to a particular amount of a pharmaceutical composition comprising a therapeutic agent that achieves a clinically beneficial result (i.e., for example, a reduction of symptoms). Toxicity and therapeutic efficacy of such compositions can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., for determining the LD50 (the dose lethal to 50% of the population) and the ED50 (the dose therapeutically effective in 50% of the population). The dose ratio between toxic and therapeutic effects is the therapeutic index, and it can be expressed as the ratio LD50/ED50. Compounds that exhibit large therapeutic indices are preferred. The data obtained from these cell culture assays and additional animal studies can be used in formulating a range of dosage for human use. The dosage of such compounds lies preferably within a range of circulating concentrations that include the ED50 with little or no toxicity. The dosage varies within this range depending upon the dosage form employed, sensitivity of the patient, and the route of administration.

The term "symptom", as used herein, refers to any subjective or objective evidence of disease or physical disturbance observed by the patient. For example, subjective evidence is usually based upon patient self-reporting and may include, but is not limited to, pain, headache, visual disturbances, nausea and/or vomiting. Alternatively, objective evidence is usually a result of medical testing including, but not limited to, body temperature, complete blood count, lipid panels, thyroid panels, blood pressure, heart rate, electrocardiogram, tissue and/or body imaging scans.

The term "disease", as used herein, refers to any impairment of the normal state of the living animal or plant body or one of its parts that interrupts or modifies the performance of the vital functions. Typically manifested by distinguishing signs and symptoms, it is usually a response to: i) environmental factors (as malnutrition, industrial hazards, or climate); ii) specific infective agents (as worms, bacteria, or viruses); iii) inherent defects of the organism (as genetic anomalies); and/or iv) combinations of these factors

The terms "reduce," "inhibit," "diminish," "suppress," "decrease," "prevent" and grammatical equivalents (including "lower," "smaller," etc.) when in reference to the expression of any symptom in an untreated subject relative to a treated subject, mean that the quantity and/or magnitude of the symptoms in the treated subject is lower than in the untreated subject by any amount that is recognized as clinically relevant by any medically trained personnel. In one embodiment, the quantity and/or magnitude of the symptoms in the treated subject is at least 10% lower than, at least 25% lower than, at least 50% lower than, at least 75%o lower than, and/or at least 90% lower than the quantity and/or magnitude of the symptoms in the untreated subject.

The term "drug" or "compound" as used herein, refers to any pharmacologically active substance capable of being administered which achieves a desired effect. Drugs or compounds can be synthetic or naturally occurring, non-peptide, proteins or peptides, oligonucleotides or nucleotides, polysaccharides or sugars.

The term "administered" or "administering", as used herein, refers to any method of providing a composition to a patient such that the composition has its intended effect on the patient. An exemplary method of administering is by a direct mechanism such as, local tissue administration (i.e., for example, extravascular placement), oral ingestion, transdermal patch, topical, inhalation, suppository etc.

The term "at risk for" as used herein, refers to a medical condition or set of medical conditions exhibited by a patient which may predispose the patient to a particular disease or affliction. For example, these conditions may result from influences that include, but are not limited to, behavioral, emotional, chemical, biochemical, or environmental influences.

The term "krill meal" as used herein, refers here to any mixture of proteins and lipids derived from krill. The term is not limited to any particular method of making krill meal, but any method known in the art is contemplated.

The term "medical disorders", as used herein, refers to any biological condition diagnosed by medically trained personell to require treatment. For example, medical disorders may include, but are not limited to, hair disorders, nail disorders, skin disorders, skeletomuscular disorders, multiple sclerosis, or sexual disorders.

The term "improved performance", as used herein, refers to any biological condition, where controlled medical testing measures results that medically trained personnel would considered above the expected norm. For example, improved performance may be measured for physical or mental tests.

The term "effective amount" refers to any amount of a supplement that improves the palatability of the food or feed.

The term "ingredient" or "supplement" refers to any composition can be formulated to a suitable form, such as a tablet, a granule, a pellet or powder. The composition may be formulated also to a pet treat or a hard gelatin capsule (sprinkle capsule) can be filled with the composition.

As used herein, the term "omega-3 fatty acid" refers to fatty acids which have the final double bond between the third and the fourth carbon atom counting from the methyl end of the carbon chain. Omega-3 fatty acids mainly concerned in this disclosure are the long chain polyunsaturated fatty acids eicosapentaenoic acid (EPA) and docospentaenoic acid (DHA) as well as the minor omega-3 fatty acids including eicosatetraenoic acid (ETA) and

docosapentaenoic acid (DPA).

As used herein, the term "lipid insoluble ingredient" refers to any compound not soluble in a lipophilic solvent such as chloroform, n-hexane and toluene. Preferably such compounds are St. John's wort, creatine, dimethyl glycine, ginko biloba, ginseng, betaglucan, mannaoligosaccharides, vincamine, whey protein, vinpocetine, zinc, zinc proteinate, copper, copper proteinate.. More preferably such compounds are grape seed extract, grape seed meal, ginger, garlic, aloe vera and green lipped mussel powder and brewer^'s yeast. Even more preferably such compounds are L-carnitine, hyaluronic acid and green tea extract. Most preferably such compounds are glucosamine hydrochloride, glucosamine sulfate, glucosamine potassium, glucosamine sodium, N-acetyl d-glucosamine, chondroitin sulfate, curcumin, ascorbic acid, manganese and manganese proteinate.

As used herein, the term "glucosamine" refers to 2-Amino-2-deoxy-D-glucose chitosamine or any derivative thereof.

As used herein, the term "chondroitin" refers to chondroitin sulfate, which is a sulfated glycosaminoglycan composed of a chain of alternating sugars (N- acetylgalactosamine and glucuronic acid) or any derivative thereof including low molecular weight forms. As used herein, the term "lipid insoluble ingredients" may also comprise any combination of the above mentioned substances. For example, one lipid insoluble ingredient comprises cartilage protecting substances such as glucosamine and/or chondroitin.

As used herein, the term "lipid insoluble vitamins" refers to vitamins or

micronutrients that are not soluble in a lipophilic solvent such as chloroform, n-hexane and toluene, but are soluble in aqueous solutions. Preferably such vitamins include, but are not limited to, vitamin B and/or vitamin C.

As used herein, the term "essential minerals" refers to minerals that are essential for the body to work and stay healthy. Preferably such essential minerals are zinc, magnesium, phosphorus, manganese, selenium, calcium, copper, iodine.

As used herein, the term "lipid insoluble ingredients" may also comprise any combination of the above mentioned substances.

As used herein, the term "lipid soluble vitamins" refers to vitamins or micronutrients that are soluble in a lipophilic solvent such as chloroform, n-hexane and toluene. Preferably such vitamins include, but are not limited to, vitamin A vitamin D, vitamin E and/or vitamin K.

As used herein, the term "bioactive health ingredients" refers to chemical compounds that are naturally occurring and that have proven biological effects or may have biological significance. Preferably such bioactive health ingredients are within the groups of carotenoids, flavonoids, isothiocyanates, phytoesterogenes, phytosterols, polyphenols. More preferably such compounds are resveratrol, beta-caroten, lycopene, lutein, zeaxantbin, genistein, curcumin, alpha-lipoic acid, ubiquinone, ubiquinol.

The term "excipients", as used herein, refer to any substance needed to formulate the composition to the desired form. For example, suitable excipients include but are not limited to, diluents or fillers, binders or granulating agents or adhesives, disintegrants, lubricants, antiadherants, glidants, wetting agents, dissolution retardants or enhancers, adsorbents, buffers, chelating agents, preservatives, colours, flavours and sweeteners. Typical excipients are for example starch, pregelatinized starch, maltodextrin, monohydrous dextrose, alginic acid, sorbitol and mannitol. In general, the excipient should be selected from non-toxic excipients (EEG, Inactive Ingredient Guide, or GRAS, Generally Regarded as safe, Handbook of Pharmaceutical Excipients). Typical excipients in particular for tableting are for example magnesium stearate, stearic acid, talc, silic, cellulose, microcrystalline cellulose, methyl cellulose, polyvinylpyrrolidone and - commercial products, such as Aerosil^®, Kollidon^® and Explotab^®. Excipients can be added into the direct powder compression formula.

The term "clinical nutrition", as used herein, refers to the study, treatment and/or prevention of nutritionally-related medical disorders, including but not limited to

malnutrition.

As used herein, the following terms are referred to in relation to krill meal use in various formulations:

- "nutritional supplement" refers to any formulation of specific ingredients that may be of short supply in routinely provided food. For example, a daily intake of a nutritional supplement is typically < 1 gram to few grams; the form of a nutritional supplement.

"food supplement" refers to any formulation of specific ingredients that may be mixed into a food to improve the food's nutritional value.

"functional food ingredient" refers to any formulation of specific ingredients that may be mixed into a food to provide an additional function, such as health-promotion or disease prevention. For example, functional foods may include, but are not limited to, processed food or foods fortified with health-promoting additives, like "vitamin-enriched" products or fermented foods with Live cultures that would confer probiotic benefits.

"food additive" refers to any formulation of specific ingredients that may be mixed into a food for purposes including, but not limited to, preservation, stability, microbial control, color, flavor etc. "pharmaceutical" refers to any formulation of specific ingredients that are used to improve the symptomology of a medical condition.

"animal feed" refers to any mixture of animal feed ingredients providing energy and nutrient requirements (e.g., protein, fat, carbohydrates, minerals and

micronutrients) in a balanced ratio. For example, a daily intake of 'animal feed' for companion animals is typically between 1 - 2% of body weight, for example 300 g/d for a 20 kg dog;

"animal feed ingredient" refers to any organic or mineral component added to an animal feed mixture in appreciable amounts (i.e., for example a bulk ingredient). Animal feed ingredients are usually added to animal feed at levels greater than 1% (w/w), preferably 1 to 90 % (w/w), typically 10 to 50 %(w/w);

"flavor ingredient" is typically an organic component added to an animal feed for the purposes of improving feed palatability. Flavor ingredients are usually added to animal feeds at levels preferably less than 1% (w/w), more preferably less than 0.5%, most preferably less than 0.1 % by weight of the animal feed;

"nutritional supplement" refers to any mixture of specific ingredients that may be of short supply in routinely provided food. For example, a daily intake of a nutritional supplement is typically < 1 gram to few grams; the form of a nutritional supplement for companion animals can be liquid or dry (e.g., pellet, tablet, granule, powder, capsule etc.);

- "treat" refers to any nutritional product form given to a companion animal as a reward or a snack between meals. Treats may contain food ingredients and other ingredients that give a special consistency to the treat e.g. chewable treat. Treats are not usually considered important for their nutritional content. Treats may be used as rewards and should be highly palatable. Alternatively, treats may also be used as sources of supplementary nutrition and should have ingredients that improve the overall nutritional balance. For example, 'healthy treats' are a special form of treats that contain specific health promoting ingredients; daily intake of a treat is typically from several grams per day up to 100 g/d for an average sized dog (20 kg body weight).

The term "fluoride" as used herein interchangeably and refer to any compound containing an organofluoride and/or an inorganic fluoride.

The term "high fluoride solid fraction" as used herein refers to a composition containing the vast majority of a crustacean's exoskeleton following a low g- force (e.g., between approximately 1,000 - 1,800 g) horizontal centrifugation separation of a hydro lyzed and disintegrated crustacean material. This fraction contains small particles of exoskeleton of the crustacean that retains the vast majority of fluoride (i.e., for example, between 50 - 95%) in these organisms.

The term "low fluoride" as used herein may refer to the product of any method and/or process that reduced the fluoride from the original material by approximately one third (i.e., for example, from 1500 ppm to 500 ppm). For example, 'a low fluoride crustacean phospholipid-protein complex' comprises one third of the fluoride luoride than 'ahydrolyzed and disintegrated crustacean material' .

The term "low fluoride hydrolyzed material fraction" as used herein refers to a composition containing the vast majority of a crustacean's fleshy internal material following a low g-force (e.g., between approximately 1,000 - 1,800 g) horizontal centrifugation separation of a hydrolyzed and disintegrated crustacean material. This fraction contains small particles of phospholipids, neutral lipids, proteins and/or peptides that is largely devoid of any fluoride (i.e., for example, between 5% - 50% of the raw hydrolyzed and disintegrated material).

The term "a low fluoride phospholipid-peptide complex composition subfraction" as used herein refers to a low fluoride composition containing the vast majority of lipid material following a high g-force (e.g., between approximately 5,000 - 10,000 g) horizontal centrifugation separation of a low fluoride hydrolyzed material fraction.

The term "concentrated hydrolysate composition subfraction" as used herein refers to a low fluoride composition containing the vast majority of water soluble peptides and/or lean material following a high g-force (e.g., between approximately 5,000 - 10,000 g) horizontal centrifuge separation of a low fluoride hydrolyzed material fraction.

The term "low fluoride oil" as used herein refers to a lipid-rich composition created by the extraction of a phospholipid-peptide complex composition subfraction using a selective extraction process, such as with a supercritical carbon dioxide fluid. Such a process removes approximately ten-fold of the fluoride from the raw hydrolyzed and disintegrated crustacean material.

The term "de-oiled phospholipid-peptide protein complex" as used herein refers to a low fluoride composition containing the vast majority of dry matter composition created by the extraction of a phospholipid-peptide protein complex composition subfraction using selective extraction process, such as a supercritical carbon dioxide fluid.

The term "phospholipid composition" as used herein refers to a low fluoride composition comprising a high percentage of polar lipids (e.g., approximately 75%) created by the extraction of a de-oiled phospholipid-peptide complex using a co-solvent, such as ethanol.

The term "extraction residue" or "protein hydrolysate" as used herein refers to a low fluoride composition comprising a high percentage of protein (e.g., approximately 60 - 90%) created by extraction of lipids from either a phospholipid-peptide protein complex or a de- oiled phospholipid-pep tide-complex using a polar solvent alone or in combination with a non-polar solvent.

The term "peptide" as used herein, refers to any of various amides that are derived from two or more amino acids by combination of the amino group of one acid with the carboxyl group of another and are usually obtained by partial hydrolysis of proteins. In general, a peptide comprises amino acids having an order of magnitude with the tens.

The term "pharmaceutically acceptable" or "pharmacologically acceptable", as used herein, refer to molecular entities and compositions that do not produce adverse, allergic, or other untoward reactions when administered to an animal or a human.

The term, "pharmaceutically acceptable carrier", as used herein, includes any and all solvents, or a dispersion medium including, but not limited to, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetable oils, coatings, isotonic and absorption delaying agents, liposome, commercially available cleansers, and the like. Supplementary bioactive ingredients also can be incorporated into such carriers.

The term, "purified" or "isolated", as used herein, may refer to a peptide composition that has been subjected to treatment (i.e., for example, fractionation) to remove various other components, and which composition substantially retains its expressed biological activity. Where the term "substantially purified" is used, this designation will refer to a composition in which the protein or peptide forms the major component of the composition, such as constituting about 50%, about 60%, about 70%, about 80%, about 90%, about 95% or more of the composition (i.e., for example, weight/weight and/or weight/volume). The term "purified to homogeneity" is used to include compositions that have been purified to 'apparent homogeneity" such that there is single protein species (i.e., for example, based upon SDS-PAGE or HPLC analysis). A purified composition is not intended to mean that some trace impurities may remain.

As used herein, the term "substantially purified" refers to amino acid sequences, that are removed from their natural environment, isolated or separated, and are at least 60% free, preferably 75% free, and more preferably 90% free from other components with whic they are naturally associated. An "isolated peptide or protein" is therefore a substantially purified peptide or protein.

Brief Description Of The Figures

Figure 1 presents a flow diagram of one embodiment of a method to produce a low fluoride crustacean material.

Figure 2 present exemplary data showing the extraction efficiencies of two different runs in accordance with one embodiment of the present invention. Detailed Description Of The Invention

The present invention is related to high quality crustacean phospholipid-peptide- protein (PPC) compositions for use as nutritional supplements, cosmetic products, pharmaceuticals and clinical nutrition. For example, the use of crustacean PPC compositions can treat medical disorders and/or to improve mental and/or physical performance. In particular, PPC may comprise a phospholipid-peptide-protein complex or a de-oiled phospholipid-pep tide-protein complex. Clinically, such crustacean PPC compositions can improve symptoms of various medical disorders.

One embodiment of the invention provides a composition comprising a crustacean meal (e.g., for example, a krill meal). In one embodiment, the crustacean meal comprises a nutritional supplement composition including, but not limited to a phospholipid-peptide complex (PPC).

The composition may further comprise excipients suitable for consumption by an animal (i.e., for example, a companion animal or a human). The composition is particularly suitable for companion animals, such as dogs or cats.

I. Krill

Antarctic krill {Euphausia Superba) is a small shrimp like crustacean which can be found in huge quantities in the Southern Ocean, off the coast of Antarctica. In order to adapt to an ever changing supply of food, krill has developed a unique survival strategy comprising highly potent digestive enzymes to facilitate complete and rapid breakdown ingested food. However, these potent digestive enzymes also result in a rapid breakdown of proteins and lipids (proteolysis and lipolysis, respectively) after harvesting of the krill. For this reason the nutrient content of krill varies, depending on harvesting area and season. In order to minimize post-harvest enzymatic activity, the krill can be frozen or turned into a meal immediately after harvesting (i.e., for example, on board the fishing vessel).

The lipids found in krill are rich in phospholipids and especially phosphatidyl choline with high content of long chain omega-3 fatty acids, in addition to the antioxidant astaxanthin. Gentle processing methods enable the levels of these nutrients to remain stable throughout the process.

Previously, a number of processes for making krill meal have been disclosed. For example krill meal can be obtained by cooking krill, followed by pressing, drying and milling, using the same technology and principles as for making fish meal. Alternatively, the krill can be cooked at two different temperatures, where the majority of the phospholipids are removed at a lower temperature. United States Patent Application Publication Number 2009/0061067, herein incorporated by reference. Other methods of making krill meal have been disclosed, where a dried powdery and granular krill product containing all components of krill was obtained. United States Patent Application Publication Number 2003/0113432, herein incorporated by reference. The production process comprised the steps of lightly dehydrating krill, coarsely crashing the krill, and drying the coarsely crushed krill under heating. Other known methods include the use of freeze drying followed by milling and the use of alcohol to denature the enzymes. Japanese Patent Publication No, 57-11876.

Alternatively, a protein rich krill meal can be obtained by enzymatic hydrolysis as previously disclosed. United States Patent Application Publication Number 2006/0035313; and WO 2010/030193, both herein incorporated by reference.

Krill contain high proportions of protein, lipid and pigments which, together with their high abundance, make them an attractive nutritional source both for animal and humans.

Experiments have shown that krill meal has a nutritional value equal to, or surpassing, that of regular fish meals when used as a substitute in the diets of various farmed species, including Atlantic cod, Atlantic salmon and Pacific white shrimp (Yoshitomi et al. 2007; Karlsen et al. 2006; Opstad et al. 2006; Gaber 2005).

Krill are believed to contain high levels of the carotenoid antioxidant astaxanthin and omega-3 fatty acid, which both ingredients is shown to have many beneficial health effects. Following death krill deteriorate very rapidly. This spoilage occurs through the rapid leaching of fluoride from the krill exoskeleton into the meat and because of auto lytic digestion by hydrolytic enzymes. Spoilage through autolysis is a major problem in krill fisheries and processing and can result in substantial losses of proteins and lipids via the generation of undesirable free fatty acids. The omega-3 polyunsaturated fatty acids present in krill oxidize quite rapidly under oxygen rich atmosphere. The astaxanthin protects the polyunsaturated fatty acid from oxidation, but as a result the astaxanhtin will be destroyed. Omega-3 polyunsaturated fatty acids in krill meal are more easily oxidized than in krill oil products and it is challenging to make stable protein-oil products.

II. Nutritional And Health Consequences Of Aging

Nutritional needs are known to change as animals (e.g., mammals, such as humans) age. Reasons for these changes include, but are not limited to, decreased absorption of essential nutrients from diet, a more sedentary lifestyle, a decreased appetite and/or a change in metabolism. These nutritional changes occur in both the healthy elderly population and the elderly with exceptional nutritional needs due to health problems or diseases. There is growing scientific documentation that shows that certain essential macro and micro nutrients can prevent the development of certain diseases in the elderly population.

There has surprisingly been found that certain novel krill meal compositions can be mixed with both Upid soluble and lipid insoluble ingredients to make stable compositions that are effective in alleviation, prevention or treatment certain diseases in the elderly population or in groups of particular nutritional needs. It has also been surprisingly found that novel krill meal compositions enhance the absorption of some essential minerals and lipid soluble health ingredients and that these krill compositions are effective in prevention and treatment of health conditions related to aging.

In one embodiment, the present invention contemplates a use of a nutritional supplement composition by administering the composition ranging between approximately 0.005 - 0.50 g krill meal per day per kg of body weight of an animal for the treatment of a degenerative joint disease. In one embodiment, the composition is homogenous. In one embodiment, the krill meal comprises a krill oil. In one embodiment, the composition further comprises at least one omega-3 fatty acid. In one embodiment, the composition is stable. In one embodiment, the homogeneity of the composition is characterized by a lack of phase separation. In one embodiment, the stability of the composition is characterized by, in comparison to krill oil, lower lyso-phospholipids, increased omega-3 levels, lower peroxide, lower p-anisidine, and higher astaxanthin.

A. Homeostatic Control

Aging may be characterized by an inability of tissues to maintain homeostasis.

This leads to an impaired response to stress and, as a consequence, an increased risk of morbidity and mortality. The incidence of numerous debilitating chronic diseases, such as cardiovascular disease, neurodegeneration, diabetes, arthritis, and osteoporosis, increases almost exponentially with age. Aging is thought to be driven, at least in part, by the accumulation of stochastic damage in cells. This includes damage to proteins, DNA, mitochondria, and telomeres, which is driven by reactive oxygen species. Mitochondria are the major producers of reactive oxygen species, which damage DNA, proteins, and lipids if not rapidly quenched. Alterations in mitochondria have been noted in aging, including decreased total volume, increased oxidative damage, and reduced oxidative capacity. These biochemical and bioenergetic changes are accompanied by perturbations in cellular dynamics, such as a decrease in mitochondrial biogenesis and an increase in mitochondrially mediated apoptosis. Courtney M. Peterson et al, Journal of Aging Research, Epub July 19, 2012.

A lack of homeostasis control can lead to an impaired response to stress and, as a consequence, an increased risk of morbidity and mortality. For example, reports suggest that the incidence of numerous debilitating chronic diseases, such as cardiovascular disease, neurodegeneration, diabetes, arthritis, and osteoporosis, increases almost exponentially with age (Tilstra, J.S. et al., The Journal of Clinical Investigation, 122(7):2601-2612 (2012)). Aging is associated with progressive loss of neuromuscular function that often leads to progressive disability and loss of independence. The term sarcopeniais now commonly used to describe the loss of skeletal muscle mass and strength that occurs in concert with biological aging. The prevalence of sarcopenia, which may be as high as 30% for those over 60 years, will increase as the percentage of the very old continues to grow in our populations. The link between sarcopenia and disability among elderly men and women highlights the need for continued research into the development of the most effective interventions to prevent or at least partially reverse sarcopenia. The aging process is also believed to be a factor in the age-dependent occurrence of central nervous system disabilities, such as dementia.

The ability of a cell to resist oxidant damage during homeostatic imbalance is determined by a balance between the generation of reactive oxygen species and the defensive capacity to produce antioxidants. Glutathione (γ-glutamylcysteinylglycine) is the most abundant endogenous intracellular antioxidant present in millimolar quantities within cells. Glutathione plays a central role in antioxidant defenses, and irreversible cell damage supervenes when the cell is unable to maintain intracellular glutathione concentrations. Evidence from several animal and human studies suggests that concentrations of glutathione decline with aging. It has been shown that dietary supplementation with the glutathione precursors cysteine and glycine fully restores glutathione synthesis and concentrations and lowers levels of oxidative stress and oxidant damages in elderly persons. Rajagopal V, Sekhar et al, Am J Clin Nutr 2011;94: 847-53. Other naturally occurring bioactive compounds, such as pyrroloquinoline quinone (PQQ), resveratrol, genistein, hydroxy-tyrosol, and quercetin have also been reported to improve mitochondrial respiratory control or stimulate mitochondrial biogenesis.

Cell permeable peptide antioxidants have been reported that are very potent at reducing intracellular ROS and preventing cell death. The peptides are tetrapeptides with alternating aromatic residues and basic amino acids. Zhao, K., et al, The Journal of

Biological Chemistry, 2004, 279, 34682.

As many of the components in krill meal might work as antioxidants and /or affect the antioxidative/inflammatory defense in the cell it would be interesting to test the krill meal and the krill meal compositions in biological systems that can measure the antioxidative effect and the anti-inflammatory effect in vitro to identify the most interesting composition. The compositions should also be tested in bioavailability studies to identify if the compounds of interest will be absorbed.

B. Age-Related Macular Degeneration

Age-related macular degeneration (AMD) is also a major cause of disability in the elderly. More than 20 million people worldwide are severely affected either by age-related macular degeneration or cataracts. AMD is the leading cause of blindness in people over 55 years of age in the western world. Nearly 30% of Americans over the age of 75 years have early signs of AMD and 7% have late stage disease. There are currently no effective treatment strategies for most patients with AMD, attention has focused on efforts to stop the progression of the disease or to prevent the damage leading to AMD.

C. Diabetes

Type 2 diabetes is the most common chronic metabolic disease in the elderly, affecting ~30 million individuals 65 years of age or older in developed countries. The estimated economic burden of diabetes in the United States is ~$ 100 billion per year, of which a substantial proportion can be attributed to persons with type 2 diabetes in the elderly age group. Epidemiological studies have shown that the transition from the normal state to overt type 2 diabetes in aging is typically characterized by a deterioration in glucose tolerance that results from impaired insulin-stimulated glucose metabolism in skeletal muscle (Petersen et al, Science, 300(5622): 1140-1142 (2003). D. Inflammation

Recent scientific studies have advanced the notion of chronic inflammation as a major risk factor underlying aging and age-related diseases. Low-grade, unresolved, molecular inflammation is described as an underlying mechanism of aging and age-related diseases, which may serve as a bridge between normal aging and age-related pathological processes.

Elevated oxidative stress has been linked to chronic inflammation and several aging related illnesses. The ability of a cell to resist oxidant damage is determined by a balance between the generation of reactive oxygen species and the defensive capacity to produce antioxidants. A central problem associated with the assessment of free radical induced oxidative stress in disease development has been the limitation in existing assay methods for in vivo measurement of free radical generation. For example, F2-isoprostanes, structural isomers of PGF2a, are formed during free-radical catalysed peroxidation of arachidonic acid. A major F2-isoprostane, 8-iso-PGF2a, is now a well-recognised reliable indicator of oxidative stress in vivo (Basu, S. Antioxid. Redox Signal. 10: 1405-1434 (2008)).

The transcription factor NF-κΒ is a component of the cellular response to damage, stress, and inflammation. Numerous studies report increased NF-kB activity with aging, and NF-kB was identified as the transcription factor most associated with mammalian aging based on patterns of gene expression (Adam et al, Genes and Development, 3244 (2007). Chronic activation of NF-κΒ is observed in numerous age-related diseases including, but not limited to, muscle atrophy, multiplesclerosis, atherosclerosis, heart disease, both type land 2 diabetes , osteoarthritis, dementia, osteoporosis, and cancer (Tilstra, J.S. et al, The Journal of Clinical Investigation, 122(7):2601-2612 (2012). NF-κΒ DNA binding is increased in skin, liver, kidney, cerebellum, cardiac muscle, and gastric mucosa of old rodents compared with that in young rodents. In addition, NF-i B was identified as the transcription factor most associated with mammalian aging, based on patterns of gene expression. Furthermore, chronic activation of NF-κΒ is observed in numerous age-related diseases, including muscle atrophy, multiple sclerosis, atherosclerosis, heart disease, both type 1 and 2 diabetes, osteoarthritis, dementia, osteoporosis, and cancer.

Several scientific publications strongly suggest that inhibitors of the IKK/NF-κΒ pathway may delay damage and extend health span in patients with accelerated aging and chronic degenerative diseases of old age.

Based on literature in the area of age related diseases, it is apparent that oxidative damage and low grade inflammation are central in the development of many of the age related diseases. The transcription factor NF-kB is a central component for the cellular response to these triggers. A scientific approach to the development of nutritional supplements and drugs for the aging population could be to focus on the development of effective antioxidants to target these central biological mechanisms. III. Treatment Of Medical Disorders With Crustacean PPC Mixtures

A. Central Nervous System Medical Disorders

The central nervous system is particularly vulnerable to oxidative insult on account of the high rate of 0₂ utilization, the relatively poor concentrations of classical antioxidants and related enzymes, and the high content of polyunsaturated lipids, the biomacromolecules most susceptible to oxidation. In addition, there are regionally high concentrations of redox-active transition metals capable of the catalytic generation of ROS. Thus, it is not surprising that oxidative stress is a common discussion point for neurodegenerative disease, where damage to neurons can reflect both an increase in oxidative processes and a decrease in antioxidant defenses.

Accumulating data indicate that oxidative stress (OS) plays a major role in the pathogenesis of multiple sclerosis (MS). Reactive oxygen species (ROS), leading to OS, generated in excess primarily by macrophages, have been implicated as mediators of demyelization and axonal damage in MS. ROS cause damage to main cellular components such as lipids, proteins and nucleic acids (e.g., R A, DNA), resulting in cell death by necrosis or apoptosis. In addition, weakened cellular antioxidant defense systems in the central nervous system (CNS) in MS, and its vulnerability to ROS effects may augmented damage. Thus, treatment with antioxidants might theoretically prevent propagation of tissue damage and improve both survival and neurological outcome. Miller, E. et al, PolMerkur Lekarski. 27(162):499-502 (2009)

B. Ocular Medical Disorders

The use of herbal medicines and nutritional supplements in ocular disorders including, but not limited to, age-related macular degeneration (AMD), cataracts, diabetic retinopathy and glaucoma, has recently been review.. Antioxidants and zinc have been used in patients with certain forms of intermediate and advanced AMD. However, there has been growing evidence regarding potential significant adverse effects associated with the AREDS (Age- Related Eye Disease Study) formula vitamins. However, whether the use of antioxidants or herbal medications in the prevention or treatment of cataracts, glaucoma or diabetic retinopathy would be beneficial is inconclusive. It was reccomended that further study of nutritional supplements and herbal medicines in the treatment of eye disease is needed to determine their safety and efficacy. Wilkinson et al., "Use of herbal medicines and nutritional supplements in ocular disorders: an evidence-based review" Drugs

24;71(18):2421-34 (2011).

Some nutritional remedies have been tried for cataracts, glaucoma, and retinal diseases (macular degeneration, diabetic retinopathy, retinopathy of the newborn, and retinitis pigmentosa). Specifically, some nutritional treatments were given for asthenopia, blepharitis, chalazion, conjunctivitis (including giant papillary conjunctivitis), gyrate atrophy of the choroid and retina, keratoconus, myopia, sicca syndrome (dry eyes), and uveitis. The data suggest that nutritional supplements may play role the further of clinical therapy strategies to ocular disorders. Gaby AR., "Nutritional therapies for ocular disorders: Part Three" Altern Med Rev. 13(3): 191-204 (2008).

C. Digestive Disorders

Functional digestive disorders can be characterized by symptoms related to the digestive tract for which no pathological causes can be found using routine diagnostic techniques. Recently, several methods have been developed to the study digestive function allow relation between in humans functional alterations, mainly motor and sensory and to be related to functional digestive symptoms. As a result of these advances, both motor and sensory alterations have been identified in subgroups of patients with functional digestive disorders. This knowledge should enable current symptom-based classifications of these disorders to be replaced with new classifications based on specific physiopathologic mechanisms. This would allow more effective therapies aimed at the specific mechanism causing the symptoms to be developed. Serra J., "Clinical research techniques in functional digestive disorders" Gastroenterol Hepatol. 29(4):255-62 (2006).

Functional dyspepsia and the irritable bowel syndrome (IBS) are amongst the most widely recognised functional gastrointestinal disorders. Symptom based diagnostic criteria have been developed and refined for the syndromes (the Rome criteria) and these are now widely applied in clinical research. Both functional dyspepsia and IBS are remarkably prevalent in the general population, affecting approximately 20% and 10% of persons, respectively. The prevalence is stable from year to year because the onset of these disorders is balanced by their disappearance in the population. Clinically useful predictors of the course of these disorders have not been identified. Approximately one third of persons with functional dyspepsia concurrently have IBS. In most studies from Western countries, it has been shown that only a minority with functional dyspepsia and IBS present for medical care; the factors that explain consultation behaviour remain inadequately defined although fear of serious disease and psychological distress may be important. The majority of patients diagnosed as having functional dyspepsia or IBS continue to have symptoms long term with a significant impact on quality of life. The indirect costs of the functional gastrointestinal disorders greatly outweigh the direct costs but overall these conditions are responsible for a major proportion of health care consumption. Rational management of the functional gastrointestinal disorders will only follow a better understanding of the natural history of these conditions. Talley N.J., "Scope of the problem of functional digestive disorders" Eur J Surg Suppl. 582:35-41 (1998).

D. Skeletal Disorders

Bone turnover, in which cells of the osteoclast lineage resorb bone and cells of the osteoblast lineage deposit bone, normally occurs in a highly regulated manner throughout life. Perturbations to these processes underlie skeletal disorders, such as osteoporosis, which are common, chronic and disabling, and increase with age. On the basis of empirical observations or on understanding of the endocrinology of the skeleton, excellent bone- resorption inhibitors, but few anabolic agents, have been developed as therapeutics for skeletal disorders. Goltzman D., "Discoveries, drugs and skeletal disorders" Nat Rev Drug Discov. 1(10): 784-796 (2002). In some embodiment, the present invention contemplates that crustacean meal compositions and other ingredients are useful in treating these disorders.

Notch signaling mediates cell-to-cell interactions that may be involved in embryonic development and tissue renewal. In the canonical signaling pathway, the Notch receptor may be cleaved following ligand binding, resulting in the release and nuclear translocation of the Notch intracellular domain (NICD). NICD induces gene expression by forming a ternary complex with the DNA binding protein CBFl/Rbp-Jk, Suppressor of Hairless, Lagl, and Mastermind-Like (Maml). Hairy Enhancer of Split (Hes) and Hes related with YRPW motif (Hey) are also Notch targets. Notch canonical signaling plays a central role in skeletal development and bone remodeling by suppressing the differentiation of skeletal cells. The skeletal phenotype of mice misexpressing Hesl phenocopies partially the effects of Notch misexpression, suggesting that Hey proteins mediate most of the skeletal effects of Notch. Dysregulation of Notch signaling is associated with diseases affecting human skeletal development, such as Alagille syndrome, brachydactyly and spondylocostal dysostosis.

Somatic mutations in Notch receptors and ligands are found in tumors of the skeletal system. Overexpression of NOTCH1 is associated with osteosarcoma, and overexpression of NOTCH3 or JAGGED 1 in breast cancer cells favors the formation of osteolytic bone metastasis. Activating mutations in NOTCH2 cause Hajdu-Cheney syndrome, which is characterized by skeletal defects and fractures, and JAGl polymorphisms, are associated with variations in bone mineral density. In conclusion, Notch is a regulator of skeletal development and bone remodeling, and abnormal Notch signaling is associated with developmental and postnatal skeletal disorders. Zanotti et al., "Notch regulation of bone development and remodeling and related skeletal disorders" Calcif Tissue Int. 90(2):69-75 (2012).

Genetic disorders involving the skeletal system may arise through disturbances in the complex processes of skeletal development, growth and homeostasis and remain a diagnostic challenge because of their variety. The Nosology and Classification of Genetic Skeletal Disorders provides an overview of recognized diagnostic entities and groups them by clinical and radiographic features and molecular pathogenesis. The aim is to provide the Genetics, Pediatrics and Radiology community with a list of recognized genetic skeletal disorders that can be of help in the diagnosis of individual cases, in the delineation of novel disorders, and in building bridges between clinicians and scientists interested in skeletal biology. In the 2010 revision, 456 conditions were included and placed in 40 groups defined by molecular, biochemical, and/or radiographic criteria. Of these conditions, 316 were associated with mutations in one or more of 226 different genes, ranging from common, recurrent mutations to "private" found in single families or individuals. Thus, the Nosology is a hybrid between a list of clinically defined disorders, waiting for molecular clarification, and an annotated database documenting the phenotypic spectrum produced by mutations in a given gene. The Nosology should be useful for the diagnosis of patients with genetic skeletal diseases, particularly in view of the information flood expected with the novel sequencing

technologies; in the delineation of clinical entities and novel disorders, by providing an overview of established nosologic entities; and for scientists looking for the clinical correlates of genes, proteins and pathways involved in skeletal biology. Warman et al., "Nosology and classification of genetic skeletal disorders: 2010 revision" Am J Med Genet A 155A(5):943- 968 (2011).

E. Muscular Disorders

Skeletal muscle is the largest organ in the human body, and plays an important role in body movement and metabolism. Skeletal muscle mass is lost in genetic disorders such as muscular dystrophy, muscle wasting and ageing. Chemicals and proteins that restore muscle mass and function are potential drugs that can improve human health and could be used in the clinic. Myostatin is a muscle-specific member of the transforming growth factor (TGF)-beta superfamily that plays an essential role in the negative regulation of muscle growth. Inhibition of myostatin activity is a promising therapeutic method for restoring muscle mass and strength. Potential inhibitors of myostatin include follistatin domain-containing proteins, myostatin propeptide, myostatin antibodies and chemical compounds. These inhibitors could be beneficial for the development of clinical drugs for the treatment of muscular disorders. Bone morphogenetic protein (BMP) plays a significant role in the development of neuromuscular architecture and its proper functions. Modulation of BMP activity could be beneficial for muscle function in muscular disorders. Tsuchida K., "The role of myostatin and bone morpho genetic proteins in muscular disorders" Expert Opin Biol Ther. 6(2): 147-154 (2006).

Currently, the diagnosis of muscular disorders is mainly clinical, wherein myopathies can present with unusual or atypical clinical features including, but not limited to, myotonia, periodic paralysis, respiratory failure, swallowing difficulties, ptosis, ophtalmoplegia, camptocormia, distal and/or asymmetrical limb muscle weakness. Several recently discovered myopathies include, but are not limited to, reducing body myopathy, X-linked myopathy with postural muscle atrophy, Emery-Dreifuss muscular dystrophy, and scapuloperoneal myopathy.

F. Cardiovascular Disorders

Dyslipidemias and insulin resistance constitute major risk factors of cardiovascular diseases (CVD) and related-features. Furthermore, oxidative stress impairment or altered antioxidant status have been suggested as pivotal keys in the onset of certain chronic diseases such as metabolic syndrome (MS), type 2 diabetes and CVD . In this sense, oxidized low- density lipoprotein (ox-LDL), a recognized oxidative stress marker, has been positively associated with central obesity, metabolic syndrome manifestations and subclinical atherosclerosis. Helen Hermana M Hermsdorff, Nutrition & Metabolism 2011, 8:59.

IV. Preparation Of Low Fluoride Crustacean Compositions

The PPC used in the methods of the present invention are created by an industrial method for processing catches of crustaceans comprising a number of steps beginning with a very early and substantially complete removal of the crustacean's exoskeleton (i.e., for example, the crust, carapace and/or shell). Although it is not necessary to understand the mechanism of an invention, it is believed that the crustacean exoskeleton comprises a vast majority of fluoride in the organism. Consequently, this step thereby results in a substantial removal of fluoride from the crustacean material. The method also uses longitudinal centrifugation techniques that prevents separation problems caused by emulsions when processing a raw material with high content of phospholipids that is initiated immediately after decking a catch of crustacean. Although it is not necessary to understand the mechanism of an invention, it is believed that the processing is initiated as soon as possible after the crustacean catch has been decked since fluoride starts to leak/diffuse immediately from the exoskeleton into the crustacean's flesh and juices. See, United States Patent Application Publication No. 2012/0149,867 (herein incorporated by reference).

When using the term "immediately" in connection with starting the process according to the present invention this relates to the period from decking the crustacean catch and to the initial disintegration of the crustacean (see infra). This period of time should be kept to a minimum, and should preferably not exceed 60 minutes, more preferred not exceed 30 minutes, even more preferred not exceed 15 minutes, and should include a direct transfer of the krill catch from the trawl bag and/or net to a suitable disintegrator. A disintegrator of the crustacean material may be a conventional pulping, milling, grinding or shredding machine. The crustacean catch is initially loaded into a disintegration appratus where the crustacean catch is subjected to pulping, milling, grinding and/or shredding to create a disintegrated crustacean material. The temperature of the disintegration process is around the ambient temperature of the water, i.e. between -2 and +1° C, preferably around +0° C. to +6° C, and may be performed by any convenient disintegration method. This disintegration process is also conventionally done by the previous known processing methods, and represents one of the obstacles according to the prior art because it produces large amounts of exoskeletal particles from the crustacean mixing in the milled material and producing a disintegrated paste with a high fluoride content. However, this high fluoride content is one of the reasons why the prior art processed crustacean material has limited applications and is less suitable for food, feed or corresponding food or feed additives compared to other marine raw materials e.g. pelagic fish.

The crustacean material may then be divided into a particle size suitable for a further separation step for not interfering with the subsequent processing steps. The disintegrating process is performed continuously and produces particle sizes up to 25 mm, a preferred particle size range is between approximately 0.5-10 mm and a more preferred size range is between approximately 1.0-8 mm.

Although it is not necessary to understand the mechanism of an invention, it is believed that this small particle size distribution represents one of advantages of the present invention because the fluoride has a tendency to leak out of the milled material and mingle with the rest of the raw material. However, this leaking process takes time and is not rapid enough to negatively impact a subsequent enzymatic hydrolysis step, provided the hydrolysis step is performed within specific parameters with respect to time and optimal, or near-optimal conditions, such as pH and temperature and optionally with the addition of co-factors such as specific ions depending on the used enzymes.

The temperature of the disintegrated material may be elevated to a temperature suitable for the subsequent enzymatic hydrolysis. Preferably, the temperature may be increased within seconds (e.g. 1-300 seconds, more preferred 1-100 seconds, even more preferred 1-60 seconds, most preferred 1-10 seconds) subsequent to the disintegrating step for reducing the processing time and thereby preventing diffusion of fluoride and for preparing the material for the enzymatic hydrolysis. Enzymes may be added directly to the disintegrated material or through the added water or both, before, during or after the disintegration process. Exogenous proteolytic enzymes (e.g., alkalase, neutrase, enzymes derived from

microorganisms including, but not limited to, Bacillus subtitis and/ 'or Aspergillus niger, and/or or enzymes derived from plant species) may be added before, during or after the disintegration, and before, during or after the heating of the disintegrated material. The added enzyme(s) may be in the form of one single enzyme or a mixture of enzymes. The conditions of the hydrolysis should match the optimal hydrolytic conditions of the added enzyme(s) and the selection of optimal conditions for the selected exogenous hydrolytic enzyme(s) is known to the person skilled in the art. As an example, the exogenous enzyme alkalase having a pH optimum of about 8, a temperature optimum of 60° C. and a hydrolysis time of 40-120 minutes. The selected enzymes, or combination of enzymes, should also be chosen for reducing emulsions caused by high content of phospholipids in the raw material.

An efficient amount of proteolytic enzyme(s) will be set after a process- and product optimization process that depends upon the efficiency of a specific chosen commercial enzyme or mix of enzymes. A typical amount by weight of commercial enzymes, as a ratio of the amount of the weight of the disintegrated raw material, are preferably between 0.5% and 0.05%, more preferably between 0.3% and 0.07% and most preferable between 0.2% and 0.09%). This hydrolysis step is aided by endogenous (natural) enzymes because rapid and uncontrolled autolysis is well known in fresh caught crustaceans.

The reason for adding exogenous enzymes is to take control of, and guide, the breakdown of the proteinaceous material in the disintegrated substance as well as speeding up/accelerating the hydrolysis of the material to avoid and/or preclude the leaking of fluoride from the shell, carapace and crust as mentioned supra. These hydrolytic enzymes, or a combination of hydrolytic enzymes, should also be carefully chosen to reduce emulsion in the production process. Enzymes may be selected from exo- and/or endopeptidases. If a mixture of enzymes is used, such a mixture may also include one or more chitinases for subsequently making the chitin-containing fraction(s) more amenable to further downstream processing. If chitinases are used, care must be taken for not increasing the leakage of fluoride from the shell/crust/carapace of the crustacean into the other fractions. However, since such fluoride leakage takes time, it is possible to perform such an enzymatic treatment within the time parameters indicated supra. A more convenient alternative to including chitinases in the enzyme mix of the initial hydrolysis step will be to process the separated chitin-containing fraction subsequently to the separation step.

As it is important to avoid the leaking of fluoride from the milled exoskeletal material into the milled fleshy material, and since the leaking to some degree is related to the increased surface area created through the disintegrating step, the enzymatic hydrolysis step should be finished within a time interval of 100 minutes, preferably within 60 minutes, most preferred within 45 minutes calculated from the addition of the endogenous enzyme(s). The amount of enzyme(s) added is related to the type of enzyme product used. As an example it may be mentioned that the enzyme alkalase may be added in an amount of 0.1-0.5% (w/w) of the raw material. This should be taken into context with the added endogenous enzymes since the addition of more enzymes will reduce the time interval of the hydro lytic step. As mentioned supra the time of the hydrolytic step is one of the crucial features of the present process since a short hydrolysis time reduces the diffusion time of fluoride from particles of the exoskeleton. The hydrolytic enzymatic processing step is intended to remove the binding between the soft tissue of the krill to the exoskeleton of the crustacean.

Subsequent to, or together with, the hydrolytic processing step the hydrolyzed and distintegraed crustacean material is passed through a particle removal device operating through a gravitational force such as a longitudinal centrifuge (i.e., for example, a decanter). This first separation step removes the fine particles containing a considerable amount of the fluoride from the hydrolysed or hydrolysing crustacean material to create a solids fraction. The centrifuge is operated with a g force between 1,000 and 1,800 g, more preferably between 1,200 and 1,600 g and most preferably between 1,300 and 1,500 g. Through this particle removal step a substantial amount of fluoride is removed from the proteinaceous crustacean fraction. The reduction of fluoride on a dry weight basis as compared to conventional crustacean meal, with a typical fluoride content of 1,500 mg/kg, maybe up to 50%, even more preferred up to 85%, most preferred up to 95%. The enzymatic hydrolysis may be terminated by heating of the hydrolysing material (incubate) to a temperature over 90° C, preferably between 92-98° C. and most preferred between 92-95° C, prior to, during or after the separation step, as long as the hydrolysis duration lies within the above given boundaries. The hydrolysis is terminated before, during, or after the fine particle removal step, most preferred after the fine particle removal step. The temperature of the first centrifugation particle removal step, in one embodiment, depend on the optimal activity temperature of the enzyme (in the case where the enzymatic hydrolysis step is terminated by heating after the fine particle separation step).

The fluoride content in the prior art processed krill protein material has limited applications and are less suitable for food or feed or corresponding food or feed additives, as mentioned supra but the fluoride content of the removed exoskeletal material is not preventive for further separation/purification of this fraction. Thus materials such as chitin, chitosan and astaxanthin may be isolated from the separated exoskeletal material. Such isolation procedures are known within the art. Steps may also be taken for removing the fluoride from the isolated exoskeletal material e.g. through dialysis, nanofiltration, through electrophoresis or other appropriate technologies.

Hydrolytic enzyme(s) deactivation may be performed in different ways, such as adding inhibitors, removing co-factors (e.g., crucial ions through dialysis), through thermal inactivation and/or by any other deactivating means. Among these, thermal inactivation, as mentioned supra, is preferred by heating the proteinaceous material to a temperature where the hydrolytic enzymes become denatured and deactivated. However, if a product where the relevant native proteins are not denatured is wanted, other means than heating for deactivating the hydrolytic enzymes should be selected.

A first centrifugation forms a de-fluorinated hydrolyzed and disintegrated crustacean material fraction and a solids fraction (e.g., containing high fluoride exoskeleton particles). As described below, the low flourine hydrolyzed and disintegrated crustacean material fraction may be subsequently separated (e.g., by a second centrifugation) to form a low fluoride Phospholipid-Peptide Complex (PPC) composition fraction, a lean low fluoride Concentrated Hydrolysate Fraction (CHF) fraction that can be used as a food and/or feed additives, and a lipid fraction mainly consisting of neutral lipids. The PPC composition subfraction is rich in lipids, like a smooth cream with no particles, wherein the lipids are well suspended within the peptide components. This suspension results in small density differences between the different PPC composition components thereby making it difficult to further separate the PPC composition with common centrifugal separators and/or decanters. This is especially accentuated with crustacean catches during the second half of the fishing season.

Ordinary disc centrifugal separators (i.e., generating rotational force in the X and Y plane) do not work properly to separate a PPC composition subfraction into its respective components since emptying and necessary cleaning cycles with water will disturb separation zones. Conventional centrifugation separation processes result in the formation of unwanted emulsion products having a high phospholipid content and low dry matter concentrations. Standard decanters cannot separate the PPC composition subfraction into its respective components due to a low g force limitation, short separation zone and an intermixing of light and heavy phases at the discharge of heavy phase from the machine.

For example, a low fluoride PPC material may be separated into subtractions using a horizontal decanter centrifuge with an extended separation path. Horizontal centrifuges (e.g., generating a rotational force in the Z plane) may comprise modified convention decanter centrifuges. For example, a PPC composition subfraction would enter an ordinary decanter from a bowl through a central placed feed pipe in the middle of the separation zone. In contrast, when using horizontal centrifuges as contemplated herein, the PPC composition subfraction enters at the end and at the opposite side of the outlet. This modification provides a significant improvement in the separation process by providing a considerably longer clarification/separation zone than ordinary decanters and utilizes the total available separation length of the machine. The drive is able to impart high g-forces: 10,000 g for small machines and 5,000 to 6,000 g for high capacity machines, facilitating the separation of very fine, slow- settling PPC composition subtractions without the complications of emulsification. The PPC composition subfraction will be subjected to the highest g-force just before entering under the baffle. The different liquid layers separated from PPC composition subfraction are concentrated gradually along the axis of the horizontal centrifuge thereby exiting the machine under baffle by the g force pressure generated by the machine. The separation of the PPC composition subfraction into a layer comprising about 27-30% dry matter makes the downstream processing efficient in terms of operating/robustness and as well economically considering both yield and costs of preparing the dry matter into a meal composition. The PPC composition subfraction separation also creates a layer comprising a lean hydrolysate that can be evaporated into a concentrated hydrolysate of greater than 60%.

In one embodiment, the present invention contemplates methods using a phospholipid- peptide complex (PPC) composition from a crustacean (i.e., for example, krill) made immediately after the catch has been brought upon on board a boat and/or ship (i.e., for example, a fishing vessel). The process of creating the PPC composition comprises disintegrating the crustaceans into a disintegrated material comprising smaller particles (i.e., for example, between approximately 1 - 25 millimeters), adding water, heating the disintegrated material, adding enzyme(s) to hydrolyze the disintegrated material, deactivating the enzyme(s), removing solids (i.e., for example, exoskeleton, shell, and/or carapace) from the enzymatically processed material to reduce the fluoride content of the material, separating and drying the PPC composition. Preferably, the PPC composition is transferred to an onshore facility (i.e., a fish oil extraction plant) where a low-fluoride crustacean oil is separated from the PPC composition using solvents including, but not limited to, supercritical C0₂ and/or ethanol. The separation of the crustacean oil from the PCC gives a high quality protein powder with a protein content of >80% with less than 10% lipids including free fatty acids. In one embodiment the present invention contemplates using this protein powder for human use in nutritional supplements or pharmaceuticals either alone or in combination with other nutrients, micronutrients or bioactive phytochemicals.

Using alternative extractions, de-oiled PPC compositions, phospolipids and/or extraction residue (i.e., for example, a protein hydrolysate) compositions are also separated from the PPC composition.

- An advantage of these crustacean products is a low fluoride content. This is due to the fact that the solid crusteacean exoskeletal particles (i.e., for example, shell and/or carapace) are effectively removed from mass to be processed.

- Another advantage that crustacean oil can be separated effectively, almost completely, from the disintegrated crustacean material (e.g., feed material) during the extraction. This is due to the fact that, in the extraction process with the supercritical C0₂ solvent, the feed material comprises a PPC composition. Although it is not necessary to understand the mechanism of an invention, it is believed that the phospholipids of the feed material are embedded in a matrix of hydrolyzed protein which means that the close association between the phospholipids and

hydrophobic/phosphorylated proteins is broken thus facilitating the extraction of the lipids.

-An advantage of this effective supercritical C0₂ extraction of the PCC is that the protein products that is remaining after separation of oil is very low in oil, free fatty acids and fluorine. In addition, the final composition is solvent-free.

-A further advantage of the PPC composition is that also the phosphatidylserine (PS), free fatty acids (FFA) and lysophosphocholine (LPC) contents are very low. V. Crustacean Phospholipid-Peptide/Protein Complexes (PPCs)

Compositions comprising crustacean lipids (e.g., for example, phospholipids, and/or omega-3 fatty acids) in the form of a dried ground meal and non-lipid ingredients overcome a number of technological problems relating to combinations of nutrients that provide a basis for in vivo synergistic effects as a result of the nutrient combination. The data provided herein demonstrate that when a crustacean PPC meal is mixed with other nutritional ingredients, the clinical improvement is unexpectedly superior to that achieved when compared to any one of the ingredients administered individually.

In some embodiments, a homogenous and stable composition is contemplated which does not separate into different phases over time. In some embodiments, the composition is formulated into a product comprising a tablet, a granule, a pellet or a powder. For example, problems of formulation are encountered when simply combining a crustacean PPC meal with other ingredients including, but not limited to, crustacean oil and/or lipid insoluble nutrients such as glucosamine, chondroitin, zinc oxide and/or vitamin C in a capsule. Simple mixing will result in a sequestration of the lipid insoluble nutrients within the capsule.

Further, the removal of sequestered nutrients from the capsule by squeezing the capsule is not possible without significant effort. The result is a situation where the daily administration of capsulated nutrients to the animal are not delivered efficiently. It is possible to combine oil and lipid insoluble nutrients also in a tablet, however, there is a limit for inclusion of oil in a tablet without extraordinary effort. United States Patent No. 5,843,919; and United States Patent Application Publication Number 2007/0113796, both herein incorporated by reference.

A. Crustacean Phospholipid-Peptide-Protein Complexes

1. Low Fluoride PPC

In one embodiment, the present invention contemplates a low fluoride phospholipid- peptide-protein complex (PPC) composition comprising proteins and peptides in the range of 40-60 weight % and lipids in the range of 40-60 weight % lipids and less than 500 mg/kg fluoride. In one embodiment, the lipids comprise phospholipids. In one embodiment, the present invention contemplates a composition comprising approximately 200-250 grams/Kg phospholipids, approximately 50-150 grams/kg Omega-3, less than 500 mg/kg fluoride, approximately 15 grams/kg lysophosphatidic acid, and less than approximately 20 grams/kg free fatty acids. The preparation of a low fluoride PPC is described herein. See, Example 1. 2. Low Fluoride Krill Meal De-Oiled Protein-Peptide Composition In one embodiment, the present invention contemplates a high quality de-oiled protein-peptide complex composition comprising approximately 90% protein/peptide, less than 500 mg/kg fluoride and 100 grams/kg lipids. The protein/peptide product results from the separation of oil from PCC. In one embodiment, the ratio between the protein fraction and the peptide fraction may range between approximately 20: 1 to 1 :20. Low Fluoride de- oiled crustacean PPC can be made as described herein. See, Example 1.

B. Crustacean PPC Mixtures

In one embodiment, the present invention contemplates a composition comprising a crustacean PPC meal and one or more additional ingredients, either alone or in combination. For example, the additional ingredient may include, but are not limited to, zinc, magnesium, calcium, vitamin C, vitamin D, vitamin E, lutein and/or zeaxanthin. In one embodiment, the composition is formulated to form a tablet, a capsule, a granule, or powder. In one embodiment, the capsule is filled with the composition. In one embodiment, the capsule is a hard gelatin capsule. In one embodiment, the hard gelatin capsule is a sprinkle capsule. Although it is not necessary to understand the mechanism of an invention, it is believed that one advantage of a sprinkle capsule is that it is easily opened. In one embodiment, a sprinkle capsule comprising krill meal and a lipid insoluble ingredient in a powder form. In one embodiment, the formulation is high in protein and lipid. In one embodiment, the formulation is low in free fatty acids. In one embodiment, the formulation is highly stable, as characterized by the low free fatty acid content, low peroxide levels, low anisidine levels and high astaxanthin levels.

1. Mixtures Of Low Fluoride PPC And Low Fluorine De-Oiled PPC a. Nutritional Supplement Compositions

In one embodiment, the present invention contemplates a nutritional supplement comprising a combination of a krill meal phospholipid-pep tide-protein complex formulation (e.g., for example, Krill Meal 1) and a second ingredient including, but not limited to, minerals, lipid soluble vitamins, lipid insoluble vitamins, bioactive health ingredients and or omega-3 oils.

In one embodiment, the present invention contemplates a nutritional supplement comprising a combination of a krill meal de-oiled phospholipid-protein complex formulation (e.g., for example, Krill Meal II) and a second ingredient including, but not limited to, minerals, lipid soluble vitamins, lipid insoluble vitamins, bioactive health ingredients and/or omega-3 oils.

Lipid Insoluble Ingredients

In one embodiment, a crustacean meal is a krill meal nutritional supplement composition comprising low fluoride krill PPC, lipid insoluble ingredients, omega-3 fatty acids, and excipients. In one embodiment, the krill PPC ranges between approximately 10 - 90 % (w/w) of the composition, more preferably ranges between approximately 20 - 60 % (w/w) of the composition, and most preferably ranges between approximately 25 to 50 % (w/w) of the composition. In one embodiment, the lipid insoluble ingredients range between approximately 10 - 90 % (w/w) of the composition, more preferably ranges between approximately 20 - 60 % (w/w) of the composition, and most preferably ranges between approximately 25 to 50 %(w/w) of the composition. In one embodiment, the omega-3 fatty acids range between approximately at least 1 % (w/w) to at least 5 % (w/w) of the composition, and preferably at least 4 % (w/w) of the composition. In one embodiment, the excipients range between approximately 0 - 80 % (w/w) of the composition, more preferably ranging between approximately 0 - 60 % (w/w) of the composition, and most preferably ranging between approximately 0 to 50 % (w/w) of the composition, wherein the composition comprises about 20 % (w/w) excipients.

In one embodiment, a daily dose of a crustacean PPC meal composition as disclosed herein is between approximately 0.1 - 100 g per day per human individual, more preferably between approximately 0.25 to 50 g/d per human individual, and still more preferably between approximately 0.5 to 25 g/d per human individual.

In one embodiment, a tablet, granule, powder, pellet or capsule comprising a crustacean PPC meal composition maybe administered in a daily dose of between approximately 0.2 to 20 g/20 kg body weight, more preferably 0.4 to 10 g/20 kg body weight, still more preferably 0.8 to 5.0 g/20 kg body weight.

In one embodiment, the present invention contemplates a composition comprising a crustacean PPC meal and lipid insoluble ingredients in a weight ratio ranging between approximately 16: 1 to 1 : 1. In one embodiment, the PPC and lipid insoluble ingredient weight ratio is approximately 4: 1. In one embodiment, the present invention contemplates a composition comprising PPC and krill oil in a weight ratio ranging between approximately 4: 1 to 1 :4. In one embodiment, the PPC and krill oil weight ratio is approximately 1: 1.

Furthermore, it is disclosed that compositions processed further into a tablet, granule, pellet, powder or treat or sprinkle capsules provide additional advantages. Inclusion of crustacean lipids in the form of PPC in tablets and other compacted products provides improved stability of crustacean lipids and this invention discloses reduced degradation of crustacean lipid components including, but not limited to, omega-3 fatty acids, astaxanthin, and phospholipids as compared to previously reported capsulated materials.

- Low Fluoride Crustacean Oil Ingredients

In one embodiment, the present invention contemplates a nutritional supplement composition, characterized in that said composition comprises a crustacean PPC meal, low fluoride crustacean oil, and lipid insoluble ingredients, hi one embodiment, the composition is homogenous. In one embodiment, the crustacean oil comprises a krill oil. In one embodiment, the composition is stable. In one embodiment, the homogeneity of the composition is characterized by a lack of phase separation. In one embodiment, the stability of the low fluoride krill oil is characterized by, in comparison to conventional krill oil, lower lyso-phospholipids, increased omega-3 levels, lower peroxide, lower p-anisidine, and higher astaxanthin.

Extracting krill oil is generally a difficult process resulting in high cost of the oil and often inferior quality (e.g., lower levels of long chain PUFA omega-3) compared to the oil in the meal. United States Patent Number 6, 020, 003; and United States Patent Number

6, 800,299, both herein incorporated by reference. In one embodiment, the present invention contemplates a crustacean lipid composition comprising at least 75% phospholipids. In one embodiment, the lipid composition comprises between approximately 75% - 90%

phospholipids. In one embodiment, the lipid composition comprises between approximately 75% - 80% phospholipids. In one embodiment, the present invention contemplates a dried extraction residue (e.g., protein hydrolysate) composition comprising approximately 70 - 80% protein, approximately 1.5 - 3.0% lipids, and approximately 5 -7 % ash. Low fluoride crusteacan oil can be made as described herein. See, Example 2.

Omega-3 Fatty Acid Ingredients

Long chain omega-3 PUFAs are one of the most studied nutrients the last 30 years and there are reported a wide variety of positive health-promoting effects and physiological functions of these omega-3 fatty acids. The two most functionally important omega-3 PUFAs are eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). Research and clinical data shown that these fatty acids are implicated in maintaining normal blood pressure, managing inflammation and supporting cognitive health. They are also central in healthy nutrition and in prevention of heart disease. EPA and DHA can be obtained from diet or produced in the human body from their precursor alpha-linolenic acid (ALA). Although conversion of v-linolenic acid to omega-3 PUFAs is important to retain constant level of EPA and DHA, emerging evidence suggest that synthesis of EPA and DHA from ALA is relative inefficient. With high age and for some diseases the body's ability to convert ALA to EPA and DHA is reduced to a level which is insufficient to retain a sufficient level in vital organs as brain and retina. Therefore, efficient tissue accretion of omega-3 PUFAs depends on the delivery of EPA and DHA from diet.

In general, the benefits of omega-3 PUFA intake are attributed to their distinct capacities to modulate cellular metabolic functions and gene expression. Variation in distribution of different fatty acids/lipids to different tissues in addition to cell specific lipid metabolism, as well as expression of fatty acid- regulated transcription factors, is likely to play an important role in determining how cells respond to changes in PUFA composition. (Benatti, P et al, J. Am. Coll. Nutr.2004, 23, 281).

The most studied omega-3 dietary source is fish oil or omega-3 concentrates from fish oil. In fish oil the omega-3 is in the form of a triglyceride and the natural content of omega-3 fatty acids is in the range of 20-30%. In concentrated formulas the omega-3 fatty acids are found as ethylesters or triglycerids and the content of omega-3 fatty acids are in the range of 50-90%. In krill oil the omega-3 fatty acids are mainly found in phospholipids and the content of omega-3 fatty acids is in the range of 15-30%.

Several studies have shown that the source and lipid form of the omega-3 have effects on absorption, distribution and tissue accumulation of the omega-3 fatty acids. This will impact the biological effects of the different omega-3 formulations. Studies have shown that polyunsaturated fatty acids given as phospholipids give increased accumulation of PUFAs in brain compared to triglycerids and ethylesters formulations.

Long chain polyunsaturated omega-3 fatty acids (omega-3 FA) present in fish oil are considered beneficial supplementary nutrients for companion animals e.g. dogs. Besides fish, omega-3 fatty acids can be derived from several sources of marine organisms including micro algae, seal, squid, molluscs, krill to name a few. Typical sources of omega-3 fatty acids from marine organisms is the oil extracted from the organism.

It is also known in the art to use meal from a marine organism containing oil as the source of omega-3 fatty acids in the animal diets. United States Patent No. 6, 054, 147, herein incorporated by reference. Supplementary nutrients are often used in combination to provide synergistic effects. For example the cartilage protecting nutrients, such as glucosamine and chondroitin, are often used in combination with omega-3 fatty acids to provide nutritional support for joints, both healthy joints and those inflicted by degenerative joint disease.

United States Patent No. 5, 843,919, WO 2007/144381, and United States Patent Application Number 2005/0101563, wherein all references are herein incorporated by reference. Lipids extracted from Antarctic krill (Euphausia superba) or krill oil contains omega-3 fatty acids.

Krill oil has been used as an ingredient in nutritional supplements with remarkable potential to alleviate the arthritis symptoms in human subjects. Deutch et al. J. Am. Coll. Nutr. 26(l):39-48 (2007). Products involving combinations of krill oil and cartilage protecting supplements, such as Genflex 3 and Omegagen are commercially available for humans. It would be useful to have similar combination products for supplementing the diets of animals, such as dogs or other companion animals suffering from osteoarthritis. The form in which the combination products are presented i.e. in gelatin capsules is not the preferred form for supplementing health promoting nutrients to dogs, as dogs do not willingly eat the capsules.

- Lipid Soluble Vitamin Ingredients

In one embodiment, the present invention contemplates a composition comprising one or more krill meal lipid insoluble ingredients, either alone or in combination. In one embodiment, the composition further comprises lipid soluble ingredients, for example vitamin A, vitamin D, vitamin E, alpha lipoic acid, lutein and other natural carotenoids. In one embodiment, the composition is formulated to form a tablet, a capsule, a granule, a pellet or powder. In one embodiment, the capsule is filled with the composition. In one embodiment, the capsule is a hard gelatin capsule.

Protein Ingredients

Food proteins have long been recognized for their nutritional and functional properties. The nutritional properties of proteins are associated with their amino acid content in conjunction with the physiological utilization of specific amino acids upon digestion and absorption. In recent years, a considerable amount of research has also focused on the liberation of bioactive peptides which are encrypted within food proteins, with a view to utilizing such peptides as functional food ingredients aimed at health maintenance. Ryan et al., Nutrients 3:765-791 (2011).

Protein is particularly important for maintaining muscle strength and preventing sarcopenia, or age-related loss of muscle mass in the elderly population. While the World Health Organization's recommended daily intake for protein is 0.8 g per kilo of bodyweight a day, many scientists working in the field suggest that 1.2 to 1.3 g/kg/day is needed to prevent sarcopenia.

Proteins from fish and other marine species have been shown to influence lipid metabolism in rodent models (Jaques et al 1995; Liaset et al., 2009) as well as reduce inflammation in response to feeding high-fat diets (Pilon et al). The combination of omega-3 fatty acids and marine protein appear to have synergistic effects on plasma lipid lowering (Wergedahl et al, 2009; Hosomi et al, 2011). For example, a fat-free krill powder comprising a hydrolysed protein demonstrated antihypertensive effects in spontaneously hypertensive rats. Specifically, two peptides with angiotensin I-converting enzyme inhibitory effects were subsequently isolated from this hydrolysed protein (Hanaka et al, 2009). A krill meal product containing lipid and protein (55 and 37%) respectively) demonstrated a broad range of effects in a mouse model of dyslipidemia and chronic inflammation. (Bjorndal et al..)

Extraction Residue Ingredients

Extraction residue (e.g., protein hydro lysate) compositions may be prepared as described herein, produced in conjunction with low fluoride de-oiled PPC. See, Example 2.

Antioxidant Ingredients

Carotenoids are phytochemicals considered beneficial in the prevention of a variety of major diseases. Carotenoids are classified, according to their chemical structure, into carotenes and xanthophylls. The carotene carotenoids include β-carotene and lycopene and the xanthophyll carotenoids include, but are not limited to, lutein, canthaxanthin, zeaxanthin, violaxanthin, capsorubin and/or astaxanthin. The marine carotenoid astaxanthin (ASTA) is naturally found in a wide variety of living organisms, such as microalgae, fungi, and crustaceans. Several studies have demonstrated that ASTA possesses powerful antioxidant properties, both in vitro and in vivo, especially as an inhibitor of LDL oxidation. Evidence has suggested that the action of carotenoids on immunity and diseases may be mediated, at least in part, by their ability to quench and/or blench ROS. In recent years, a number of studies on astaxanthin have demonstrated its in vitro and in vivo antioxidant effect, for example, the quenching effect on singlet oxygen, a strong scavenging effect on superoxide, hydrogen peroxide, and hydroxyl radicals and an inhibitory effect on lipid peroxidation. The specific molecular mechanisms of its actions are not yet established. Speranza L, Mar Drugs 10(4):890-899 (2012)

The human retina accumulates lutein and zeaxanthin two other carotenoids. The latter predominates at the macula lutea while lutein predominates elsewhere in the retina. There is epidemiological evidence of a relationship between low plasma concentrations of lutein and zeaxanthin, and an increased risk of developing age-related macular degeneration (AMD). Some studies support the view that supplemental lutein and/or zeaxanthin help protect against AMD.

There are indications that orally ingested antioxidants can be effective in lowering oxidative stress and to prevent or improve health conditions related to oxidative stress. It is important that these antioxidants have the capability to penetrate cell membranes. To have effects in relation to neurodegenerative and eye diseases the antioxidant has to penetrate the blood brain barrier. One antioxidant that has this capability is astaxanthin. Astaxanthin provides cell membranes with potent protection against free radical or other oxidative attack. Experimental studies confirm that this nutrient has a large capacity to neutralize free radical or other oxidant activity in the nonpolar ("hydrophobic") zones of phospholipid aggregates, as well as along their polar (hydrophilic) boundary zones. Kidd, P. Alternative Med. Rev. 2011, 11, 364.

Resveratrol Ingredients

Resveratrol, is a natural product isolated from most red wines. In recent years, resveratrol derivatives (including its oligomers) have shown diversity in regards to its chemical and biological activities. For example, these compounds are believed to have antioxidant activity. Such antioxidant activity of resveratrol derivatives may be dependent upon specific structure-activity relationships as evidenced by comparison of resveratrol derivatives (i.e., for example resveratrol oligomers). Others sugggest that resveratrol may also have a potential as therapeutic agents for cerebral and cardiovascular diseases. He et al., "From Resveratrol to Its Derivatives: New Sources of Natural Antioxidant" Curr Med Chem. E-pub Dec 3, 2012.

Ubiquinone Ingredients

Coenzyme Q10 (CoQIO) widely occurs in organisms and tissues, and is produced and used as both a drug and dietary supplement. Increasing evidence of health benefits of orally administered CoQIO are leading to daily consumption in larger amounts, and this increase justifies research and risk assessment to evaluate the safety. A large number of clinical trials have been conducted using a range of CoQIO doses. Reports of nausea and other adverse gastrointestinal effects of CoQIO cannot be causally related to the active ingredient because there is no dose-response relationship: the adverse effects are no more common at daily intakes of 1200 mg than at a 60 mg. Systematic evaluation of the research designs and data do not provide a basis for risk assessment and the usual safe upper level of intake (UL) derived from it unless the newer methods described as the observed safe level (OSL) or highest observed intake (HOI) are utilized. The OSL risk assessment method indicates that the evidence of safety is strong at intakes up to 1200 mg/day, and this level is identified as the OSL. Much higher levels have been tested without adverse effects and may be safe, but the data for intakes above 1200 mg/day are not sufficient for a confident conclusion of safety. Hathcock et al., "Risk assessment for coenzyme Q10 (Ubiquinone)" Regul Toxicol

Pharmacol. 45(3):282-8 (2006).

Mineral Ingredients

Although minerals are essential for all aspect of health, they tend to be poorly absorbed from diet, and the absorption becomes even less efficient in proportion with incresed age.

Zinc

Zinc was recognized to be essential for human health in 1963, and its deficiency affects nearly 2 billion people in the developing world. Growth retardation, immune disorders, and cognitive impairment are major manifestation of zinc deficiency. Prasad et al, J. Lab. Clin. Med, 138(4):250-256 (2001).

Calcium

Calcium is a mineral believed essential for living organisms, in particular in cell physiology, where movement of the calcium ion (Ca ⁺) into and out of the cytoplasm functions as a signal for many cellular processes. As a major material used in mineralization of bone, teeth and shells, calcium is the most abundant metal by mass in many animals. Calcium supplements are used to prevent and to treat calcium deficiencies. It is currently recommended that supplements be taken with food and that no more than 600 mg should be taken at a time because the percent of calcium absorbed decreases as the amount of calcium in the supplement increases. It is also recommended to spread doses throughout the day. Recommended daily calcium intake for adults ranges from 1000 to 1500 mg. It is recommended to take supplements with food to aid in absorption. Vitamin D is added to some calcium supplements. Proper vitamin D status is important because vitamin D is converted to a hormone in the body, which then induces the synthesis of intestinal proteins responsible for calcium absorption.

Disorders of calcium are usually linked to magnesium balance and, consequently, are physiologically and clinically challenging. For example, a physiology-based approach to the disorders of hypocalcemia, hypercalcemia and/or hypomagnesemia would suggest that the balance of both minerals are involved. Calcium and, to a lesser extent, magnesium balance is achieved through a complex interplay between the parathyroid gland, bone, the intestine and the kidney. Currently, the understanding of molecular physiology of calcium and magnesium balance suggests the involvement of proteins including, but not limited to, the calcium- sensing receptor (CaSR) and the main intestinal and renal transporters for calcium and magnesium, namely, the transient receptor potential channels TRPV5, TRPV6 and TRPM6. The regulation of parathyroid hormone (PTH) secretion by CaSR and the subsequent effects of PTH and vitamin D on TRPV5 constitute an increasingly characterized regulatory loop. Hoom et al., "Disorders of calcium and magnesium balance: a physiology-based approach" Pediair Nephrol. E-pub. Nov 10, 2012.

C. Veterinary Supplements

In one embodiment, the present invention contemplates a composition comprising an animal food and a crustacean PPC meal food ingredient. In one embodiment, the PPC meal food ingredient is less than 5% of the composition.

In one embodiment, the krill meal composition comprises a pet treat. In one embodiment, the pet treat composition includes flavor ingredients. Although it is not necessary to understand the mechanism of an invention, it is believed that such flavor ingredients give the pet treat composition a special appealing flavour. In one embodiment, the flavor ingredients are either natural or synthetic. In one embodiment, the pet treat compositions further comprise filler ingredients. Although it is not necessary to understand the mechanism of an invention, it is believed that such filler ingredients provide a desirable consistency and bulk to the composition. In one embodiment, filler ingredients include but are not limited to, meat meals and extracts, other animal byproducts, fish meal and extracts, plant protein meals, cereal meals or fractions of cereals and various binding agents known in the art.

In one embodiment, a daily dose of a krill meal composition as disclosed herein is administered as between approximately 0.005 to 0.50 gram (g) krill meal/kilogram (kg) animal body weight, more preferably 0.01 to 0.25 g krill meal/kg animal body weight, and still more preferably 0.02 to 0.125 g krill meal/kg animal body weight. For example, when administering a krill meal composition to a dog of 20 kg body weight, a daily dose of the composition preferably would be between approximately 0.1 - 10 g krill meal per day per animal, more preferably between approximately 0.2 to 5 g/d/animal, still more preferably between approximately 0.4 to 2.5 g/d/animal.

As described herein, animals (e.g., companion animals) prefer to consume food which contains added krill meal over food that does not contain krill meal. Companion animals have been shown to demonstrate preference to food that contains very low levels of krill meal as compared to food that does not contain krill meal. Although it is not necessary to understand the mechanism of an invention, it is believed that krill meal enhances the palatability of animal food or feed, in particular a companion animal's food or feed (i.e., for example, dog food) at concentrations that are at least an order of magnitude lower than what would be required if krill meal was added to food for its nutritional content (protein, fat, carbohydrate).

In one embodiment, the present invention contemplates a method for improving the palatability of animal food or feed, characterized in that at least 0.01 % (w/w), but less than 1 % (w/w) krill meal is added to animal food or feed.

In one embodiment, the present invention contemplates a use of krill meal as a flavor ingredient in the amount of less than 1 % (w/w) in animal food or feed.

In one embodiment, the present invention contemplates a composition comprising an animal food and a krill meal food ingredient. In one embodiment, the krill meal food ingredient comprises a krill oil. In one embodiment, the composition further comprises at least one omega-3 fatty acid. In one embodiment, the krill meal food ingredient is less than 5% of said composition. In one embodiment, the krill meal food ingredient is less than 1% of said composition, hi one embodiment, the hard gelatin capsule is a sprinkle capsule.

Although it is not necessary to understand the mechanism of an invention, it is believed that one advantage of a sprinkle capsule is that it is easily opened. In one embodiment, a sprinkle capsule comprising krill meal and a lipid insoluble ingredient in a powder form. Hard gelatin capsules are especially suitable as they can be opened easily by the companion animal owner and the contents dispensed to the animal, for example by sprinkling the composition on dog food.

It is well known in the prior art that capsulated material is difficult to administer to an uncooperative animal, in particular if the capsule is an unpalatable gelatin capsule. This invention provides a solution to this problem as it is shown that the novel compositions described herein are more willingly consumed by animals.

VI. Methods Of Producing Crustacean Meal Containing Products

In one embodiment, the present invention contemplates a method for preparing a nutritional supplement composition, characterized in that the method comprises mixing a crustacean PPC meal and lipid insoluble ingredient(s). In one embodiment, the mixing further comprises at least one excipient suitable for food or feed. A. Methods To Produce Krill Meal Nutritional Supplements

In one embodiment, the present invention contemplates a method for producing a nutritional supplement composition. In one embodiment, the method comprises mixing krill meal and lipid insoluble ingredient(s). In one embodiment, the method further comprises mixing suitable excipients. The composition is preferably formulated to a desired form by using methods well known for a person skilled in the art.

In one embodiment, the mixing produces a homogenous composition comprising dry krill meal and lipid insoluble ingredients. In one embodiment, the mixing of the dry krill meal includes krill oil. One advantage of the presently disclosed method is that, unlike previous metliods producing unstable krill oil products (since some components, in particular phospholopids may degrade with time), the krill meal krill oil as produced herein is stable.

B. Methods To Produce Palatable Animal Food

In one embodiment, the present invention contemplates a novel method of enhancing the palatability of animal food or feed, in particular dog food by adding low levels of krill meal. Furthermore, the method further comprises increasing the food intake of an animal. Although it is not necessary to understand the mechanism of an invention, it is believed that for some animals, in particular companion animals, such as finicky dogs which commonly do not eat sufficient amounts of food offered, krill meal food supplements may improved food consumption.

In one embodiment, the method further comprises adding the krill meal composition to an animal food or feed. In one embodiment, the amount of krill meal composition added to the animal food or feed is less than 5 %, preferably less than 3 %, more preferably less than 1% (w/w), still more preferably less than 0.5%, most preferably less than 0.1% of the weight of the feed. In one embodiment, the amount of krill meal composition added to the animal food or feed ranges between approximately 0.01 - 0.9 % (w/w), more preferably ranging between approximately 0.1 - 0.5 % (w/w).

In one embodiment, a krill meal food or feed comprises a highly palatable treat used for training of the dogs or other companion animals, where krill meal is used as an animal feed ingredient.

It is a surprising feature of the presently contemplated invention that very low doses of krill meal improve food and/or feed palatability. Previously reported data on the effect of krill meal as a feed attractant has been contradictory. For example, it has been observed that krill meal can be used as a feed attractant in fish, while other studies report reduced feed intake in chicken fed a diet supplemented with krill meal. United States Patent Application Publication No. 2008/0274203; and Olsen et ah, Aquaculture Nutr. 12(4):280-290 (2006); both herein incorporated by reference.

C. Methods To Produce Crustacean Meal Nutritional Supplements

In one embodiment, the present invention contemplates a nutritional supplement composition comprising krill meal lipids and lipid insoluble components. In one

embodiment, the nutritional supplement composition comprises a companion animal treat. In one embodiment, the nutritional supplement composition comprises a suitable form selected from the group consisting of a tablet, a capsule, a granule, a pellet, or a powder. In one embodiment, the capsule comprises a hard gelatin capsule. In one embodiment, the hard gelatin capsule is a sprinkle capsule. Although it is not necessary to understand the mechanism of an invention, it is believed that the nutritional supplement is a health promoting composition. Some advantages of the presently contemplated nutritional supplements includes, but is not limited to: i) that the combination of nutrients has synergistic effects; ii) that the nutritional supplements within the krill meal are stable; and iii) the nutritional supplement is easy to administer to human or non-human animal.

In one embodiment, the present invention contemplates a krill meal nutritional supplement comprising cartilage protecting substances. In one embodiment, the cartilage protecting substance comprises chondroitin and/or glucosamine. In one embodiment, the nutritional supplement may further comprise a lipid insoluble ingredient.

The invention is not limited to any particular lipid insoluble ingredient, but a range of lipid insoluble ingredients are contemplated. Non-limiting examples of such components are: glucosamine hydrochloride, glucosamine sulfate, glucosamine potassium, glucosamine sodium, N-acetyl d-glucosamine, chondroitin, hyaluronic acid, green lipped mussel powder, creatine, L-carnitine, ascorbic acid, manganese, manganese proteinate, zinc, zinc proteinate, copper, copper proteinate, ginseng, green tea extract, ginger, garlic, vincamine, grape seed extract, grape seed meal, dimethyl glycine, whey protein, brewer's yeast, St. John's wort, vinpocetine, aloe vera, ginko biloba, curcumm, betaglucans, mannaoligosaccharides and any combination thereof.

The invention is not limited to the particular methodology, protocols, and reagents described herein because they may vary. Further, the terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of the present invention. VII. Pharmaceutical Compositions And Methods Of Making

The present invention further provides pharmaceutical compositions (e.g., comprising the compounds described above). The pharmaceutical compositions of the present invention may be administered in a number of ways depending upon whether local or systemic treatment is desired and upon the area to be treated. Administration may be topical

(including ophthalmic and to mucous membranes including vaginal and rectal delivery), pulmonary (e.g., by inhalation or insufflation of powders or aerosols, including by nebulizer; intratracheal, intranasal, epidermal and transdermal), oral or parenteral. Parenteral administration includes intravenous, intraarterial, subcutaneous, intraperitoneal or intramuscular injection or infusion; or intracranial, e.g., intrathecal or intraventricular, administration.

Pharmaceutical compositions and formulations for topical administration may include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids and powders. Conventional pharmaceutical carriers, aqueous, powder or oily bases, thickeners and the like may be necessary or desirable.

Compositions and formulations for oral administration include powders or granules, suspensions or solutions in water or non-aqueous media, capsules, sachets or tablets.

Thickeners, flavoring agents, diluents, emulsifiers, dispersing aids or binders may be desirable.

Compositions and formulations for parenteral, intrathecal or intraventricular administration may include sterile aqueous solutions that may also contain buffers, diluents and other suitable additives such as, but not limited to, penetration enhancers, carrier compounds and other pharmaceutically acceptable carriers or excipients.

Pharmaceutical compositions of the present invention include, but are not limited to, solutions, emulsions, and liposome-containing formulations. These compositions maybe generated from a variety of components that include, but are not limited to, preformed liquids, self-emulsifying solids and self-emulsifying semisolids.

The pharmaceutical formulations of the present invention, which may conveniently be presented in unit dosage form, may be prepared according to conventional techniques well known in the pharmaceutical industry. Such techniques include the step of bringing into association the active ingredients with the pharmaceutical carrier(s) or excipient(s). In general the formulations are prepared by uniformly and intimately bringing into association the active ingredients with liquid carriers or finely divided solid carriers or both, and then, if necessary, shaping the product.

The compositions of the present invention may be formulated into any of many possible dosage forms such as, but not limited to, tablets, capsules, liquid syrups, soft gels, suppositories, and enemas. The compositions of the present invention may also be formulated as suspensions in aqueous, non-aqueous or mixed media. Aqueous suspensions may further contain substances that increase the viscosity of the suspension including, for example, sodium carboxymethylcellulose, sorbitol and/or dextran. The suspension may also contain stabilizers.

In one embodiment of the present invention the pharmaceutical compositions may be formulated and used as foams. Pharmaceutical foams include formulations such as, but not hmited to, emulsions, microemulsions, creams, jellies and liposomes. While basically similar in nature these formulations vary in the components and the consistency of the final product.

Agents that enhance uptake of oligonucleotides at the cellular level may also be added to the pharmaceutical and other compositions of the present invention. For example, cationic lipids, such as lipofectin (U.S. Pat. No. 5,705,188), cationic glycerol derivatives, and polycationic molecules, such as polylysine (WO 97/30731), also enhance the cellular uptake of oligonucleotides.

The compositions of the present invention may additionally contain other adjunct components conventionally found in pharmaceutical compositions. Thus, for example, the compositions may contain additional, compatible, pharmaceutically-active materials such as, for example, antipruritics, astringents, local anesthetics or anti-inflammatory agents, or may contain additional materials useful in physically formulating various dosage forms of the compositions of the present invention, such as dyes, flavoring agents, preservatives, antioxidants, opacifiers, thickening agents and stabilizers. However, such materials, when added, should not unduly interfere with the biological activities of the components of the compositions of the present invention. The formulations can be sterilized and, if desired, mixed with auxiliary agents, e.g., lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, colorings, flavorings and/or aromatic substances and the like which do not deleteriously interact with the nucleic acid(s) of the formulation.

Dosing is dependent on severity and responsiveness of the disease state to be treated, with the course of treatment lasting from several days to several months, or until a cure is effected or a diminution of the disease state is achieved. Optimal dosing schedules can be calculated from measurements of drug accumulation in the body of the patient. The administering physician can easily determine optimum dosages, dosing methodologies and repetition rates. Optimum dosages may vary depending on the relative potency of individual oligonucleotides, and can generally be estimated based on EC50S found to be effective in in vitro and in vivo animal models or based on the examples described herein. In general, dosage is from 0.01 μg to 100 g per kg of body weight, and may be given once or more daily, weekly, monthly or yearly. The treating physician can estimate repetition rates for dosing based on measured residence times and concentrations of the drug in bodily fluids or tissues. Following successful treatment, it may be desirable to have the subject undergo maintenance therapy to prevent the recurrence of the disease state, wherein the compound is administered in maintenance doses, ranging from 0.01 μg to 100 g per kg of body weight, once or more daily, to once every 20 years.

Experimental

Example 1

Preparation Of Low Fluoride Crustacean Meals

The preparation of low fluoride crusteacean meal (e.g., krill) is discussed herein in greater detail (supra). See, Figure 1. A fresh crustacean catch underwent disintegration and hydrolysis immediately after decking onboard a fishing vessel. Subsequently, the crusteacean exoskeleton was separated from the fleshy proteinaceous/lipid material to form a low fluoride phospholipid-peptide-protein complex (low fluoride PPC), wherein the majority of the fluoride in the organism is retained within the exoskeleton material (e.g., by using a horizontal centrifuge). This low fluuride fleshy proteinaceous/lipid material was then dried and ground into a low fluoride crustacean meal (e.g., for example, Krill Meal 1). An overall analysis of Krill Meal 1 is depicted in Table 1 :

Table 1 : Overall Composition Analysis

Lipids 45 g/lOOg

Fluoride 400 ppm

Astaxanthin 300 ppm

Further analysis provided a further composition breakdown of Krill Meal 1 as determined by standard methods in the art and the results are shown in Tables 2 - 5 below.

Table 2. Fatty Acid Composition

Fatty acids mg/g

Sum saturated 221

Sum monounsaturated 145

Sum total PUFA 323

Sum total PUFA n-3 294

Sum total PUFA n-6 2,2

EPA + DHA 221

EPA/DHA 1,4

Table 3. Lipid Class Composition

Lipid classes g/100g

TG 36

DG 0,8

MG <1

FFA 1,8

Sterols 1,9

Pho sphatidylcholine 44

Total neutral 40

Total polar 53,2

Table 4. Peptide Molecular Weight Distribution of water soluble peptides

Molecular weight of peptide % of peptides

>70000 2,2

70000-60000 0,3

60000-50000 0,2 50000-40000 0,2

40000-30000 0,2

30000-20000 0,4

20000-10000 1,0

10000-5000 8,0

5000-1000 11,1

<1000 76,3

Table 5. Amino Acid Content

Krill Meal 1, or a low fluoride PPC, was further subjected to an extraction with a supercritical gas (e.g., carbon dioxide) that created a low fluoride de-oiled phospholipid- protien complex (e.g., low fluoride de-oiled PPC). The low fluoride de-oiled PPC was then dried and ground into Krill Meal 2. An overall analysis of Krill Meal 2 is depicted in Table Table 6: Overall Composition Analysis

Table 9. Peptide Molecular Weight Distribution

Example 2

Production Of Low Fluoride Krill Oil

A feed material, such as low fluoride PPC or Krill Meal 1 was prepared in accordance with Example 1, was supplied in a sealed plastic bag containing approximately 25kg. The feed material was kept frozen until used in extractions. The granules have a size distribution typically in the range 2 to 5mm, but a number of fine fragments were also present. The granules are greasy to the touch but still break up under compression rather than smear.

5 kg batches of feed material in granular form, as processed using supercritical C(¾ as solvent and azeotropic food grade ethanol as co-solvent, the weight of the ethanol being 23% of the weight of C0₂. The plant was pre-pressurised to operating pressure with C0₂ only, and ethanol was added when C0₂ circulation started. Solvent to feed material ratio was 25: 1 or greater and co-solvent to feed material ratio was 5:1. Runs were carried out under two extraction conditions; 300 bar at 60°C, and 177 bar at 40°C. See, Table 10.

Table 10 -Krill Oil Extraction Conditions

Run 1 Run 2

Feed Mass (g, as received) 5000.5 5000.9

Extraction pressure (bar) 300 177 Extraction temperature (°C) 60 33

First separator pressure (bar) 90 90

First separator temperature (°C) 41 41

Second separator pressure (bar) 48-50 48-50

Second separator temperature (°C) 39 39

C0₂ used with ethanol co-solvent (kg) 132.6 134.9

Additional C0₂ at end of run (kg) 33.1 44.5

Total ethanol used (kg) 31.65 32.19 The extracted krill oil material was passed through two separation vessels in series, held at 90 bar and 45-50 bar respectively. The final krill oil material collected from both separators was pooled together and the ethanol was evaporated. The residual feed material comprises a de- oiled feed material (e.g., for example, de-oiled PPC) and/or an extraction residue, such as a protein hydrolysate, having a reduced lipid content in comparison to the starting feed material. See, Example IX.

After ethanol evaporation, krill oil cumulative extraction curves were generated for both Run 1 and Run 2 by independently analyzing each sample taken during the extraction runs. See, Table 11. Table 11 - Progressive krill oil extraction sample points and yields.

Sample Number 1 2 3 4 5 6 Total Run 1

Cumulative C0₂ (kg/kg feed) 5.5 9.1 13.4 17.8 22.0 33.1 33.1

Extracted oil (g, dry) 1137 398 282 135 78 86 2115

Run 2

Cumulative C0₂ (kg/leg feed) 5.6 9.1 13.5 17.5 21.5 34.4 34.4

Extracted oil (g, dry) 715 496 368 220 149 129 2077 A total yield of 41-42 wt% of the feed material was achieved for all runs. The runs carried out at 300 bar and 60°C had a higher initial rate of extraction. The curves indicate that the extraction is virtually complete at Sample Number 5 after a cumulative C0₂ use ranging between 21.5 - 22.0 kg per kg of feed material. Estimated maximum extraction is achieved at a point where the C0₂:feed ratio is 26.5: 1. See, Figure 3 (estimated maximum extraction is marked by an arrow). The ratio of azeotropic ethanol to C0₂ was 0.24: 1 for the 300 bar runs, and slightly higher at 0.26: 1 for the lower pressure run.

This method of krill oil production resulted in the near complete extraction of total lipids from the krill meal (e.g., for example, approximately 95% of neutral lipids and 90% of phospholipids. The final yield was similar for both the high and low pressure runs, but neutral lipids were more rapidly extracted at higher pressure. The phospholipid extraction rate was similar under both extraction conditions. As detailed below, in this extraction process, the pooled krill oil total lipid had an overall phospholipid level of just over 40 wt% and both phosphatidyl inositol and phosphatidyl serine were poorly extracted.

Phospholipid profiles of the various krill material compositions were then determined using traditional column chromatography techniques. See, Table 12.

Table 12 - Comparative Phospholipid Profiles Of Krill Compositions (run 1)

The first column shows the specific phospolipids that were analyzed. The second column show the phospholipid profile of the starting feed material (e.g., a low fluoride PPC prepared as described in Example 1). Columns three - eight (Extracts 1 - 6) show the phospholipid profile of each krill oil sample taken during the extraction process as described above. The last two columns show the phospholipid profile of the residual extracted feed material sampled from either the top and/or the bottom of the phospholipid extraction column (e.g., for example, an extraction residue, protein hydrolysate and/or de-oiled PPC).

The data show that the major phospholipid in the extracted krill oil samples is phosphatidyl choline (PC), ranging approximately from 72.7% to 80.4% of total

phospholipids, including contributions from both alkyl acyl phosphatidyl choline (AAPC) and lyso phosphatidyl cholines (e.g., for example, LPC and/or LAAPC). Smaller amounts of phosphatidyl ethanolamine (PE) are present in both the feed material (column 1, - 5.3%) and in the krill oil extract samples (columns 3 - 8), ~ 3.5 - 4.5%). Alkyl acyl and lyso forms of PE (AAPE, LPE) are also present in the feed material and krill oil extracts. Phosphatidyl inositol (PI) and phosphatidyl serine (PS) are present in the feed material, but because they are poorly soluble in ethanol, these phospholipids are poorly extracted and are therefore concentrated in the extracted feed material residue (e.g., having a higher level in the residual PPC in comparison to the feed material, see columns 9 and 10).

Further analysis determined the overall relative lipid component proportions of the extracted krill oil. See, Table 13.

Table 13 - Main Lipid Components Of Extracted Krill Oil (%w/w)

The data show: i) a relative absence of free fatty acids (FFAs); ii) less than 2% of sterols; iii)

40 wt% of triacylglycerides (TAGs); and iv) approxiately 50% phospholipids (e.g., polar lipids). While FFA's were not detected (ND) in this particular example, it is believed that extracted krill oils may comprise between approximately 0.01 - 0.1 % FFA of total lipids.

As described above, the extraction process results a yield of between approximately 92.2 -

95.3% of the feed material total lipid.

The method and products according to the invention has been described above. The method can naturally vary in its details from those presented. The inventive idea may be applied in different ways within the limits as described herein.

Example 3

Lipid Extraction Efficiency

This example demonstrates an exemplary analytical lipid extraction with the Soxhlet method comparing conventional krill meal with a low fluoride krill meal (e.g. low fluoride PPC) as described herein. Soxhlet method is a standard method in quantitative determination of fat content of foods and feeds and thus it can be used as a reference method to determine the extractability of various krill meals. For example, the Soxhlet method may be carried out as below using petroleum ether (boiling point 30-60 °C). Conventional krill meal was prepared as described in US 2008/0274203 (Aker Biomarine ASA, Bruheim et al.) and the low fluoride PPC was prepared according to the present invention.

The neutral lipids are often part of large aggregates in storage tissues, from which they are relatively easily extracted. The polar lipids, on the other hand, are present as constituents of membranes, where they occur in a close association with proteins and polysaccharides, with which they interact, and therefore are not extracted so readily. Furthermore, the phospholipids are relatively tightly bound with hydrophobic proteins and in particular with the phosphorylated proteins.

The data show that partial hydrolysis of the protein matrix in the preparation of a low fluoride PPC composition as described herein improves the extraction efficiency of total lipid by use of non-polar organic solvents (e.g., for example, supercritical C0₂, ethanol, and/or petroleum ether).

Briefly, a 10 g sample of either conventional milled krill meal or low fluoride PPC was weighed and placed in a Soxhlet apparatus and then continuously extracted for approximately eight (8) hours using 300 mL petroleum ether. After extraction, the solvent was evaporated at 60 °C under a nitrogen stream. Soxhlet F., "Die gewichtsanalytische bestimmung des milchfettes" Dingier 's Polytech. J. 232:461-465 (1879).

The results show that the proportion of residual (e.g., un-extracted) lipid was twice as large in the conventional krill meal compared to the low fluoride krill meal. See, Table 14.

Table 14: Lipid Extraction Efficiency Of Low Fluoride Krill Meals

Consequently, the lipid extraction methods described herem have provided an unpredictable and surprising result that provides a superior product because of a greatly improved extraction efficiency.

Example 4

Determination Of Fluoride Content

This example presents one method of determining fluoride content of krill products as fluoride by chemical analysis using an ion selective electrode.

A low fluoride PPC (Krill Meal 1) was prepared as described herein and extracted in accordance with Example 2 to create a low fluoride krill oil. Both the meals and the oils were analyzed for fluoride content and compared with conventional preparation processes. Briefly, the method disclosed herein removes, in most part, the krill exoskeleton from the krill meal thereby reducing the fluoride content. In contrast, the krill exoskeleton is included in the conventional krill meal thereby having relatively high levels of fluoride. Conventional processes are, for example, described in WO 2002/102394 (Neptune Technologies &

Bioresources) and US 2008/0274203 (Alter Biomarine ASA).

The krill meals analyzed for fluoride content were produced by: i) a low fluoride method of present invention; and ii) a whole krill material produced by a conventional process. See, Table 15.

Table 15: Fluoride Content Comparison To Conventional Processes

The data demonstrate that by removing the exoskeleton in the process of producing krill meal (e.g., the low fluoride preparation as disclosed herein), the fluoride content of the krill meal and the krill oil produced from the meal have a markedly reduced fluoride content (e.g., 3 - 10 fold reduction). Example 5

Krill Oil Color Comparison

Krill oil has typically a strong red colour arising from the carotenoid astaxanthin present in the oil at levels varying from 50 ppm to 1500 ppm. Color of krill oil can be determined with a LabScan^® XE spectrophotometer (Hunter Associates Laboratory, INC. Resbon, VA, USA) and reported in CIELAB colour scales (L*, a* and b* values). Deviation from the red colour of astaxanthin can occur when the krill biomass is processed at high temperature and under conditions that induce oxidation. Typical oxidation induced deviation in krill oil color is an increase in the brownish hue. Brown color in krill oil arises from oxidation of lipids and formation of secondary and tertiary oxidation products with amino residues. This process is also called non-enzymatic browning.

Strecker degradation products and pyrroles are products of non-enzymatic browning that have been characterized in samples of krill oil. For example, polymerization of pyrroles results in formation of brown, melatonin like macromolecules. Furthermore, pyrrole content of krill oil can be determined spectroscopically with absorbance at 570 nm.

Samples of three krill oils will be examined for color. One produced by the method of the present invention, one produced from frozen krill by a method described in WO 2002/102394 (Neptune Technologies & Bioresources) and one extracted from dried krill meal with ethanol alone as described in US 2008/0274203 (Aker Biomarine ASA). It is to be found that krill oil produced by the method of the present invention has the lowest level of brown color determined spectrophotometrically by using CIELAB colour scales (L*, a* and b* values) and/or the lowest level of pyrroles determined spectroscopically.

Example 6

Organoleptic Krill Oil Quality Determination

Organoleptic quality of krill oil is conventionally determined by chemical analysis of volatile nitrogenous compounds arising from the decomposition of krill proteins and trimethyl amine oxide (TMAO). Nitrogenous compounds analyzed are total volatile nitrogen (TVN) and trimethylamine (TMA). In simplified terms the level of nitrogenous compounds correlate with the level of spoilage in the raw material i.e. krill biomass used for extraction of the oil.

It has become evident that, in addition to the volatile nitrogenous compounds, a large number of volatile components with distinct odour contribute to the sensory properties of laill oil. Many of the volatile components arise from the oxidation of lipid and proteinaceous compounds of krill biomass. Thus, a method that limits the level of oxidative degradation in the krill biomass, will reduce the amount of volatile components in krill oil.

Assessment of the organoleptic quality of different types of krill oil is to be performed by a panel of trained individuals. The sensory properties to be determined include several pre-defined parameters of smell and taste. It is to be found that the novel krill oil has an improved sensory profile compared to the other oils tested. The other oils to be tested include one extracted from frozen krill by a method described in WO 2002/102394 (Neptune Technologies & Bioresources) and one extracted from dried krill meal with ethanol alone as described in US 2008/0274203 (Aker Biomarine ASA).

Example 7

Production Of Low Trimethyl Amine Crustacean Products This example describes one method to produce low TMA crustacean products using a krill meal material composition. One having ordinary skill in the art, upon reading this specification would understand that this krill meal material composition may have variable fluoride content, including fluoride contents below 0.5 ppm, in addition to the basic components described below. See, Table 16. Table 16: Unextracted Krill Meal Composition

Alternatively, krill oil was prepared by krill meal extraction at 40 bars and 40°C using supercritical dimethyl ether (SC DME). The DME extract composition was dried on a Rotavapor^® and then flushed with nitrogen. The components of the resultant dried composition is listed below. See, Table 18.

Table 18: Krill Oil Components After SC DME Extraction Of Krill Meal

These data clearly show that supercritical DME extraction of krill meal compositions result in a preferential 10 - 100 fold reduction of TMA and TMAO levels. Example 8

Nuclear Magnetic Resonance Phospholipid Profiles Of Low Fluoride Krill Oil This example presents representative data of the phospholipid composition of low fluoride krill oils prepared by the methods described herein. See, Table 19.

Table 19: Phospholipids in Low fluoride krill oil analyzed using P NMR.

Sample #1 (color; orange)

Sample #2 (color; orange)

Sample #3 (color; orange)

sampte sodds

Sim of the Identified phaspteipid

Sample #4 (color; orange)

** May contain 8«n<¾ PO) Sample #5 (color; orange)

These data are consistent with those obtained using traditional column chromatography techniques shown in Example I.

Example 9

Lipid Compositional Analysis Of Low Fluoride PPC Material The example presents data showing the lipid compositional analysis of a low fluoride phospholipid-protein complex composition created by the methods described herein.

Consequently, it would be expected that the fluoride content of the compositions described below are less than 500 ppm.

The PPC comprises approximately 46.7 g/100 g (e.g., ~ 47%) total fat, 11.8 g/100 g (e.g., ~ 12%) eicosapentaenoic Acid (EPA) and 6.7 g/ 100 g (e.g., ~7%) docosahexaenoic acid (DHA). The total lipid content of the PPC total fat was approximately 87.7 % (w/w) and comprises between approximately 115 - 260 mg/kg astaxanthin and between approximately

35.2% - 46.7% unextracted oil. Table 20: Low Fluoride Krill PPC Fat: Neutral Lipid Content (45.2% w/w of total fat): Sample Number IMG

Table 21 : Low Fluoride Krill PPC Fat: Neutral Lipid Content (46.6% w/w of total fat):

Sample Number 2MG

Table 22: Low Fluoride Krill PPC Neutral Lipids: Fatty Acid Content (49.7% w/w of neutral lipids): Sample Number IMG

Table 23: Low Fluoride Krill PPC Neutral Lipids: Fatty Acid Content (46.7% w/w of neutral lipid): Sample Number 2MG

Example 10

Lipid Compositional Analysis Of Low Fluoride De-Oiled PPC Material (Krill Meal 2) The example presents data showing the lipid compositional analysis of a low fluoride de-oiled phospholipid-protein complex composition created by the methods described herein. Consequently, it would be expected that the fluoride content of the compositions described below are less than 500 ppm. The de-oiled PPC comprises approximately 35 g/ 100 g (e.g., ~ 35%) total fat, 16.6 g/100 g (e.g., ~ 17%) eicosapentaenoic Acid (EPA) and 10.0 g/ 100 g (e.g., -10%) docosahexaenoic acid (DHA). The total lipid content of the de-oiled PPC total fat was approximately 87.7 % (w/w) and comprises approximately 115 mg/kg astaxanthin and approximately 35.2% unextracted oil. Table 26 : Low Fluoride Krill De-Oiled PPC Fat: Neutral Lipid Content (20.1 % w/w of total

Table 27: Low Fluoride Krill De-Oiled PPC Neutral Lipids: Fatty Acid Content (35.2% w/w

Table 28: Low Fluoride Krill PPC De-Oiled Polar Lipid Content (68.9% w/w of total fat): Sample Number 3MG

Example 11

Compositional Analysis Of PPC/Extraction Residue Mixtures The example presents data showing the lipid compositional analysis of a low fluoride phospholipid-protein complex mixed with an extraction residue (protein hydrolysate) composition created by the methods described herein in an approximate 60/40 ratio.Protein was prepared by extraction of lipids from either the PPC or the de-oiled PPC. It would be expected that the fluoride content of the compositions described below are less than 500 ppm. The mixture comprises between approximately 28-30 g/100 g (e.g., ~ 30%) total fat, approximately 98 mg/kg astaxantine esters, approximately less than 1 mg/kg astaxanthine, a peroxide level of less than 0.1 %;(mEq/kg) and/or an ananiside level of less than 0.1 % (w/w).

Table 29: Low Fluoride PPC/Protein Mixture Fat: Neutral Lipid Content (28% w/w of total fat)

Table 31: Low Fluoride PPC/Protein Mixture Polar Lipid Content

Example 12

Composition 3

Krill meal 1 was mixed with zinc oxide, Vitamin C and vitamin E, wherein the daily effective dose is between approximately 1 - 3 grams. This composition is effective for ocular medical disorders, such as those involving retinal disorders.

Example 13

Composition 4

Krill meal 1 is mixed with zinc oxide, vitamin C, vitamin E, lutein, and zeastaxanthin, wherein the daily effective dose is between approximately 0.5 - 2 grams. This composition is effective for ocular medical disorders, such as those involving retinal disorders.

Example 14

Composition 4

Krill meal 1 was mixed with phytosterols and vitamin D, wherein the daily effective dose is between approximately 0.5 - 4 grams. This composition is effective for cardiovascular medical disorders, such as those involving the heart.

Example 15

Composition 5

Krill meal 1 mixed with calcium, vitamin D and vitamin K, wherein the daily effective dose is approximately between 0.5 - 4 grams. This composition is effective for skeletal medical disorders, such as those involving overall bone health.

Example 16

Composition 6

Krill meal 1 was mixed with Ubiquinone (Q10). The composition is effective for central nervous system disorders, such as those involving the brain.

Example 17

Composition 7

Krill meal 2 was mixed with microencapsulated DHA, lutein, xeastazantin, zinc, and vitamin C. This composition is effective for ocular medical disorders, such as those involving the retina.

Example 18

Composition 8

Krill meal 2 was mixed with microencapusulated omega-3 fatty acids. This composition is effective for muscular medical disorders, such as those involving muscle protein anabolism and/or mitochondrial function. Dysfunctional mitochondria in particular are thought to play a key role in muscle function decline, as the mitochondria are the main producers of both cellular energy and free radicals. Alterations in mitochondria have been noted in aging, including decreased total volume, increased oxidative damage, and reduced oxidative capacity.

Example 19

Preparation Of Krill Meal Hard Gelatin Capsules

Hard gelatin capsules are commercially available (e.g., PAG Pharmatech Co., Ltd. SZ China (Mainland)) and generally have two components a base, or body, and a shorter cap which fits firmly over the base. A variety of capsules sizes are available, wherein the capacity of each size varies according to PPC composition and their apparent densities. Hard gelatin capsules are commercially available as clear gelatin capsules or in a variety of colors. For example, different colored capsules can be used to distinguish two different PPC formulations thereby allowing easy identification. The capsule halves are separated and a predetermined, weighed amount, of PPC composition is placed in each capsule half. The capsule halves are then inserted into each other and stored until use.

Example 20

Stability Of A Krill Meal 1 in Hard Gelatine Capsules

Hard gelatine capsules are filled with a PPC composition in accordance with Example 1 . Different subsets of the filled capsules are then stored under different environmental conditions, for example, at temperatures ranging between - 10 to +100 ° C for different lengths of time (e.g., ranging between 24 hours - 5 years). It would be expected that little or no changes in PPC composition occurs at the lower temperature and shorter lengths of time, versus d e higher temperatures for longer periods of time.

Example 21

Preparation Of Krill Meal Composition In Flavored Drinks

The mixtures of crustacean PPC composition as described herein are used to flavor drinks and/or beverages. Such drinks and/or beverages include, but are not limited to, sport drinks, nutritional drinks, milk drinks (e.g., cow milk, goat milk etc.). The flavoring may be performed on an individual drink were a predetermined, weighed amount, of PPC composition is mixed into the drink. Alternative, on a commercial scale, the flavoring may be performed on a large volume of the drink, (e.g., tens, hundreds or thousands of gallons) by emulsifing the PPC composition into the drink volume. Example 22

Total Antioxidant Capacity Measurements

Total antioxidant capacity is measured in different crustacean low fluoride PPC compositions using a Ferric Reducing Antioxidant Power (FRAP) analysis (Vitas AS).

Total antioxidant activity is measured by a previously reported ferric reducing antioxidant power (FRAP) assay. Benzie et al., "Ferric Reducing/ Antioxidant Power Assay: DirectMeasure of Total Antioxidant Activity of Biological Fluids and Modified Version for Simultaneous Measurement of Total Antioxidant Power and Ascorbic Acid Concentration" Methods In Enzymology 299:15-23 (1999). FRAP assay uses antioxidants as reductants in a redox-linked colorimetric method, employing an easily reduced oxidant system presentin stoichiometric excess.

At low pH, reduction of ferric tripyridyl triazine (Fe III TPTZ) complex to ferrous form (which has an intense blue colour) can be monitored by measuring the change inabsorption at 593 nm. The reaction is non specific, in that any half reaction that has lower redox potential, under reaction conditions, than that of ferric ferrous half reaction, will drive the ferrous (Fe III to Fe II) ion formation. The change in absorbance is therefore, directly related to the combined or "total" reducing power of the electrondonating antioxidants present in the reaction mixture. A list of primary reagents are:

a) Acetate buffer 300 mM pH 3.6

b) TPTZ (2, 4, 6-tripyridyl- s - triazine) (M.W. 312.34) 10 mM in 40mM HC1 c) FeCl₃-6H₂0

d) Ascorbic Acid (M.W. 176.13) 1000 μ M

A sample (100 μΐ) of a PPC composition is mixed with 3 ml of working FRAP reagent and absorbance (593 nm) is measured at 0 minute after vortexing. Thereafter, samples are placed at 37°0 C in water bath and absorption is again measured after 4 minutes. Ascorbic acid standards (100 μΜ-1000 μΜ) were processed in the same way. The analyzer or spectrophotometer is blanked and the OD of Standard and Test at zero minute is measured at 593 nm.

A FRAP value of the sample (μΜ) = (Change in absorbance of sample from 0 to 4 minute / Change in absorbance of standard from 0 to 4 minute) X FRAP value of standard (1000 μΜ). Example 23

Inhibition of NF- Β by Krill Meal 1

Krill Meal 1 is prepared in accordance with Example I, flushed with argon and stored at 0°C until use.

Cell cultures:

U937-NE-KB-LUC cells, are cultured in RPMI-1640 medium with L-glutamin (2nM), penicillin (50 U/ml), streptomycin (50 mg/ml), hygromycin (75 ug/ml), 10% Fetal Bovine Serum at 37°C and C0₂. The cells are seeded in 24-well plates wherein 1% Fetal bovin serum is added to the medium. NF-kB activity is induced by lipopolysaccharide (LPS) or human TNF-a. Cell viability was measured by trypan blue staining.

Luciferase activity assay:

Luciferase activity is measured by imaging with a IVIS Imaging system from

Xenogen Corp., USA. The Luminescence is detected after 1 min. and 5 min after addition of 0.2 mg d-luciferin/ml cell medium. Number of photons in each well/second is calculated using Living Image Software. (Xenogen)

Results:

Some of the PPC compositions as described herein will show potent inhibition of the

NF-kB pathway.

Example 24

In Vivo Testing

Mouse Models

To test the effect of a crustacean PPC composition on lipid parameters and atherosclerosis parameters, the preferred animal model could be either the ApoE-3*Leiden mice or the ApoE-3 *Leiden.CETP mice. The lipid profile of the mice model is similar to humans and the animal respond to omega-3 pufas, sterols, statins. Measurements include, but are not limited to, standard lipid parameters as TG, LDL, HDL but also include some cardiovascular inflammatory parameters as ox-LDL, LP-PLA2 and CRP which are relevant as biomarkers for atherosclerosis. It it is expected that crustacean PPC meal has a greater therapeutic efficacy than ordinary omega-3 products on these parameters.

A relevant animal model for inflammation is the NFKB-RE-1UC transgenic mouse model. This animal model express luciferase under the control of NF-kappaB, enabling real- time in vivo imaging of NF-kappaB activity and thereby inflammation in intact animals.

Human Clinical studies:

Clinical studies with crustacean PPC nutraceuticals/nutritional supplements as described herein are administered to a population of elderly patients. The data show a reduction in at least one age-related biomarker thereby preventing the development of an age- related health disorder.

For example, one age-related biomarker comprises an F2-isoprostane. F2-isoprostanes are related a group of bioactive prostaglandin F2-like compounds generated by oxidatively catalyzed reaction of arachidonic acid, are considered as the reliable marker of lipid peroxidation in vivo. The 8-isoprostane (8-isoprostaglandin F2a; the major F2-isoprostane), the well- known compound belonging to the F2-isoprostane class, is usually quantified in urine instead of plasma for practical use because of the short half- life of plasma F2- isoprostane. Elevated levels of plasma and/or urinary 8-isoprostane have been reported in several conditions such as diabetes , alcoholic liver disease, and cardiovascular disease.

Clinical studies are also performed by administering crustacean PPC composistions are described herein to patient populations with risk of developing a medical disorder selected from the group comprising macular degeneration (e.g., for example, development of drusen), atherosclerosis (biomarkers may include, but are not limited to, CRP, oxidized LDL, isoprotanes) and osteoporosis.

These in vivo models (or similar ones known to those having ordinary skill in the art) can be used to determine: i) a superior reduction in drusen spots in retina by treatment with Krill Meal 1 or Krill Meal 2, and the additional ingredients of zinc, xeastazantin, lutein and astaxanthin; ii) an improved lipid profile and reduction of inflammatory markers in APOE3 mice with either Krill Meal 1 or Composition 4 supplemented with krill oil and 60% omega-3 fattu acids; iii) a reduction of mflammatory markers in humans with cardiovascular risk factors CRP, Oxidized LDL, isoprotanes, TG, LDL, HDL. Example 25

Comparative Dietary Absorption Of A PPC Composition And

Commercially Available Astaxanthin Compositions

The absorption of astaxanthin from a PPC composition as described herein is compared with a commercially available astaxanthin supplement product.

HPLC atmospheric pressure chemical ionization (APCI)/MS, GC MS, HPLC diode array detection (DAD), and NMR is used for the identification of astaxanthin and astaxanthin fatty acid esters in krill (Euphausia superb Dana). Matrix solid phase dispersion is applied for the extraction of the carotenoids. This gentle and expeditious extraction technique for solid and viscous samples leads to distinct higher enrichment rates than the conventional liquid-liquid extraction. The chromatographic separation is achieved employing a C30 RP column that allows the separation of shape-constrained geometrical isomers.

A methanol/tert-butylmethyl ether/water gradient maybe applied. Astaxanthin and the geometrical isomers are identified by HPLC APCI/MS, by coelution with isomerized authentical standard, by UV spectroscopy (DAD), and three isomers are unambiguously assigned by microcoil NMR spectroscopy. In this method, microcoils are transversally aligned to the magnetic field and have an increased sensitivity compared to the conventional double-saddle Helmholtz coils, thus enabling the measurement on small samples. The carotenol fatty acid esters are saponified enzymatically with Lipase type VII from Candida rugosa. The fatty acids are detected by GC MS after transesterification, but also without previous derivatization by HPLC APCI/MS. C14:0, C16:0, C16: l, C18: l, C20:0, C20:5, and C22:6 are found in astaxanthin monoesters and in astaxanthin diesters. Astaxanthin is identified as the main isomer in six fatty acid ester fractions by NMR. Quantitation is carried out by the method of internal standard. (13-cis) Astaxanthin (70 microg/g), 542 microg/g (all- E) astaxanthin, 36 microg/g unidentified astaxanthin isomer, 62 microg/g (9-cis) astaxanthin, and 7842 microg/g astaxanthin fatty acid esters are found.

The data is expected to show that absorption of PPC compositions is greater than commerically available products. Example 26

Comparative Dietary Absorption Of A PPC Composition And

Commercially Available Calcium Compositions

The absorption of calcium carbonate is compared between a commercial calcium dietary supplement with Composition 5. The assay is performed using a calcium switch assay with Caco-2 cells.

Cell culture

Caco-2, a human colonic epithelial cell line, are obtained from the American Type Culture Collection and maintained routinely in 100- x 15-mm petri dishes at 37°C with 5% C0₂ 95% air atmosphere and >95% humidity.

The cell monolayers are grown in DMEM (Mediatech) supplemented with 10% fetal bovine serum, 100 kiu/L penicillin, 100 mg/L streptomycin, 1-glutamine (2 mmol/L), and non-essential amino acids. All experiments are performed in the same medium without 10% fetal bovine serum, i.e. in the serum-free medium.

Calcium switch assay

Fully confluent Caco-2 cell monolayers (usually within 72 h after confluence) are washed with calcium-free GIBCO Minimum Essential Medium (S-MEM) and treated with S- MEM for i 6 h.

S-MEM is removed and cell monolayers are washed 3 times with serum-free DMEM. The cell monolayers are then incubated in the regular serum-free DMEM for pre-specified time periods under standard cell culture conditions. Reassembly of tight junctions and restoration of barrier function are determined at various time points by measuring the TER.

The data is expected to show that a PPC composition calcium source show superior calcium availablity (i.e., faster switching) that commercially available calcium supplements.

Example 27

Stability Of A PPC Composition Supplement Tablet

A PPC composition is made according to Example I but is mixed with glucosamine, chondroitin and a natural antioxidant (for example green tea extract or tocopherols) or other natural health promoting ingredients (for example curcumin). Additional ingredients are mixed with the PPC composition to include a ratio of krill oil omega-3 fatty acid to glucosamine and chondroitin matches with what is considered to be beneficial to alleviate the symptoms in dogs with osteoarthritis. For example, PPC composition 30 %, glucosamine and chondroitin 30 % and excipients 30 % (w/w) are mixed together. The stability and homogeneity of the product is studied.

The product is further processed into a tablet using known methods in the art. In the tablet there is at least 4 % krill oil omega-3 fatty acids. A tablet and a prior art soft gelatin capsule are administered to a dog and compared, whether the dog prefers to consume the tablet over the capsule or vice versa.

Example 28

Stability Of A Commerically Available Krill Meal Nutritional Supplement Tablet QRILL^® krill meal was obtained from Aker Biomarine (Oslo, Norway) and mixed with glucosamine, chondroitin, a natural antioxidant (vitamin-C) and curcumin.

The ingredients were mixed so that the ratio of krill oil omega-3 fatty acid to glucosamine and chondroitin matched with what is considered to be beneficial to alleviate the symptoms in dogs with osteoarthritis.

Here, krill meal 30 %, glucosamine and chondroitin 30 % and excipients 30 % (w/w) were mixed together and processed into a tablet using known methods in the art. The product was found to be homogenous with respect to the oil content, but it was not stable with respect to the oil quality. The level of free fatty acids and lyso-phospholipids increased gradually over time.

A tablet and a prior art soft gelatin capsule filled with krill oil were administered individually to a group of ten dogs. It was found that 9 out of 10 dogs ate the tablets, none of the dogs ate the capsule voluntarily.

Example 29

Comparative Stability Of Krill Meal Tablets Versus Krill Oil A krill oil product is obtained (Neptune Biotech, Laval, Canada) and combined with commercially available glucosamine and chondroitin into a liquid product. The chemical stability of the three products is studied and compared.

After a certain amount of time, the levels of lyso-phospholipids, omega-3 fatty acids, peroxide value, p-anisidine value andastaxanthin in the tablets are compared with their levels in the liquid krill oil product. Krill oil and glucosamine + chondroitin are mixed in ratios 4:1, 1 : 1 and 1 :4 and it is studied whether the lipid (krill oil) and non-lipid (glucosamine + chondroitin) phase are separated. In addition, krill meal and glucosamine + chondroitin are mixed in ratios 4: 1, 1 :1 and 1 :4 based on the oil content of krill meal and a tablet using methods known in the art is produced.

The homogeneity and stability of the krill meal and glucosamine + chondroitin tablets is compared to the homogeneity and stability of the capsulated krill oil glucosamine + chondroitin mixture.

It is to be observed that the liquid krill oil products result on a phase separation and is therefore not homogenous. It is also to be observed that the liquid krill oil product compared to the tablets results in higher levels of lyso-phospholipids, lower omega-3 levels, higher peroxide values, higher p-anisidine values and lower Astaxanthin values. These results clearly show that there is an improved chemical and physical stability of the krill tablets compared to the krill oil product.

Example 30

Palatability Of Krill Meal Nutritional Supplements

Tablets made with PPC compositions are prepared in accordance with Example I were tested for palatability with 10 dogs. The control tablet contains only glucosamine and chondroitin as active ingredients.

Palatability tests are performed as follows: each dog receives three products: i) Krill Meal 1 tablets; ii) Krill Meal 2 tablets); and iii) control tablets. Tablets are administered daily with food in the morning at 1 tablet/10 kg body weight for 30 days. The order of administration of these tablets is random, but is recorded with observations on palatability.

No difference is found in palatability between Krill Meal 1 and Krill Meal 2, but the Control is clearly less palatable. Often the Control tablet remained in the bowl after the food is consumed.

Example 31

Palatability Of Commercially Available Krill Meal Food Additives QRILL^® Krill meal is obtained from Aker Biomarine (Oslo, Norway) and added to dry animal food at four different levels of inclusion: 5%, 1%, or 0.1% (w/w) level and one formulation without any krill meal (0% or Control).

These four foods are offered to a group of dogs to determine the relative palatability of foods with lirill meal versus Control. The palatability test is performed as a 'two-bowl-test' where each of the dogs are offered one bowl of food with krill meal (5, 1 or 0.1%) and one bowl of food without krill meal (Control). The amount of food in each of the bowls is roughly 50% of the daily allowance of the dog. Dogs are allowed to eat until one of the bowls is empty or until both of the bowls are half empty. The preference is recorded and 'no preference' is recorded in case the dogs ate from both bowls.

Γη most cases the foods with krill meal is more palatable. There was no clear effect of the inclusion level. Roughly 20% of dogs did not show preference, but ate from both bowls. Very few dogs preferred the Control food suggesting that adding krill meal to the food enhanced the palatability of the food.

Example 32

Palatability Of Fresh PPC Composition Food Additives

PPC compositions made in accordance with Example I is added to dry animal food at four different levels of inclusion: 5%, 1%, or 0.1% (w/w) level and one formulation without any PPC (0% or Control).

These four foods are offered to a group of dogs to determine the relative palatability of foods with krill meal versus Control. The palatability test is performed as a 'two -bowl-test' where each of the dogs are offered one bowl of food with PPC (5, 1 or 0.1%) and one bowl of food without krill meal (Control). The amount of food in each of the bowls was roughly 50% of the daily allowance of the dog. Dogs are allowed to eat until one of the bowls is empty or until both of the bowls are half empty. The preference is recorded and 'no preference' is recorded in case the dogs ate from both bowls.

In most cases the foods with PPC is more palatable. There is no clear effect of the inclusion level. Roughly 20% of dogs did not show preference, but ate from both bowls. Very few dogs preferred the Control food suggesting that adding PPC to the food enhanced the palatability of the food.

Example 33

Preparation Of Healthy Companion Animal Treats

A healthy treat comprising a PPC composition made in accordance with Example I and cartilage protecting substances is prepared. The formula of healthy treat combines 2000 mg of PPC, 500 mg of glucosamine + chondroitin and 2500 mg of base ingredients for a total weight of 5 g/treat. Base ingredients include, but are not limited to, meat meals and extracts, other animal by-products, fish meal and extracts, plant protein meals, cereal meals or fractions of cereals and various binding agents known in the art. Pet treats may also contain additional ingredients that provide desirable consistency and bulk to the product. Alternatively, the 5 g treat can be produced from PPC and cartilage protecting substances alone by mixing 4500 mg of PPC + binding agents with 500 mg of glucosamine + chondroitin. After mixing the ingredients the mixture is pelleted, extruded or otherwise processed into treats of different shapes.

Example 34

Preparation Of Krill Meal Hard Gelatin Capsules

The mixture of ingredients described in accordance with Example 1 are used to fill hard gelatin capsules (e.g., sprinkle capsules) in accordance with Example 19. The capsule is opened and its contents sprinkled over a bowl of food. The palatability of the capsules is assessed and compared with control capsules in accordance with Example 32.

Example 35

PPC Composition Nutritional Supplement Treatment of Degenerative Joint Disease

A nutritional supplement is prepared for dogs comprising a PPC composition made in accordance with Example 1 and cartilage protecting substances.. The supplement combined 90% of PPC and 10% glucosamine hydrochloride + chondroitin sulfate (in 60:40 ratio). After mixing, the ingredients are pelleted with a matrix type granulation unit and stored in aluminum foil plastic bags under a nitrogen atmosphere. This supplement is then used in a clinical test involving 10 dogs with degenerative joint disease (i.e., for example,

osteoarthritis; OA).

Ten dogs of various ages and breeds presenting with different types of joint problems resulting in various degrees of OA are recruited to participate in a 6 week test. OA is often associated with pain and thus it influences the movement of the dog. Before starting the administration of the supplement, the dogs are brought to the veterinary clinic for examination which includes, but is not limited to, weighing, body condition scoring, palpation of the joints and observations on the movement of the dogs.

A force plate (4LegCheck; ReDog, Vasteras, Sweden) is used to determine the distribution of weight between the four legs while the dog was standing. Deviation from equal distribution of weight (± 5%) between the left and right leg suggests pain/lameness. Also, deviation from the typically observed 60:40 distribution of weight (± 5%) between front and rear legs is considered to be a sign of lameness. The dogs are then fed their normal diet and nutritional supplement (5 g/lOkg body weight) for six weeks. After six weeks, the clinical examination is repeated as described above. In addition to the clinical re-examination, the dogs' owner fills in a questionnaire regarding the mobility of the dog by using a modified Helsinki Chronic Pain Index (HCPI) questionnaire (Hielm- Bjorkman et al. 2003) with 11 questions relating to movement. The scores range from 1 to 5 where 1 represents greatly reduced mobility, 3 represents no change and 5 represents greatly improved mobility. The mean score of the 11 questions is used as the mobility index. Thus, three types of observations regarding the change in dogs mobility are performed: i) observations made by the clinician; ii) assessment of the owner performed by using a validated questionnaire; and iii) a device providing highly repeatable measurements on lameness based on weight distribution between the four legs of the dog.

The data show a consistent improvement in the mobility of the dogs during the six week nutritional supplementation period. Briefly, force plate measurements indicate that 7 out of 10 dogs demonstrated reduced lameness due to supplementation. Also, owner assessment show that the average mobility score improved in 8 out of 10 cases with mobility indexes ranging from 3.5 to 5. In one dog there is no change, and in one dog the changes are both positive and negative resulting hi overall score of 3. This dog demonstrated significant improvement in mood, activity and playing. However, increased mobility is associated with greatly increased difficulty in moving after rest. Finally, the clinical examination by the veterinarian documented 6 cases of improved movement, 5 cases of reduced pain and one case of reduced lameness and pain, respectively. In total, 8 out of 10 dogs demonstrated improvement in one or more clinical endpoints.

It is of particular interest to examine the supplementation of the dogs prior to the test. One dog had been supplemented with a combination of omega-3 fish oil and a joint health product (Seraquin by Boehringer Ingelheim), four dogs had been supplemented with a joint health product (Cartivet by Biofarm and Arthroflex by ScanVet) and five dogs had received no supplementation. Seraquin, Cartivet and Arthroflex are products that contain glucosamine and chondroitin as the main chondroprotective nutrients. All five dogs receiving nutritional supplements prior to the test start respond with significant improvement in joint health and mobility suggesting that the combination of krill meal and chondroprotective nutrients (glucosamme + chondroitin) as in the current composition and applied during the 6 week test period is more effective than fish oil + chondroprotective nutrients (Seraquin) or chondroprotective ingredients alone (Cartivet, Arthroflex). Example 36

PPC Composition Treatment of Degenerative Joint Disease A nutritional supplement for dogs comprising a PPC composition is prepared having the ingredients described in accordance with Example 1. PPC is then pelleted with a matrix type granulation unit and stored in aluminum foil plastic bags under nitrogen atmosphere. Pelleted PPC is used as a supplement in a clinical test involving 8 dogs with degenerative joint disease (i.e., for example, osteoarthritis; OA). The test was performed by a veterinary clinic.

Eight dogs of various ages and breeds presenting with different types of joint problems resulting in various degrees of OA are recruited to participate in a 6 week test.

Before starting the administration of the supplement the dogs were brought to the veterinary clinic for examination which includes a clinical examination, measurements on a force plate and owner assessment of the mobility. Supplement was fed to the dogs at 5 g/10 kg of body weight. All three measurements of pain and mobility provide evidence of improvement. However, the overall improvement is somewhat less than what is observed for the combination of PPC and glucosamine + chondroitin.

Briefly, force plate measurements demonstrate significant reduction in lameness in 2 out of 8 dogs. Owner's assess an average mobility score improved in 5 out of 8 cases with mobility indexes ranging from 3.25 to 4.5. In 3 out of 8 cases there was no change. Finally, the clinical examination by a veterinarian documents 2 cases of improved movement, 1 case of reduced pain, 1 case of reduced lameness and 2 cases of reduced stiffness. In total, 4 out of 8 dogs demonstrate improvement in one or more clinical endpoints. In this group the dogs demonstrate, in general, less symptoms at the baseline and their forceplate measurements also indicate less lameness. Thus, the apparent lower efficacy of the PPC supplement compared to the combination supplement may be influenced by the random selection of less symptomatic test subjects into this group.

It is also of interest to examine the supplementation of the dogs prior to the test with krill meal alone. One dog had been supplemented with a combination of omega-3 fish oil and a joint health product (Seraquin by Boehringer Ingelheim), four dogs had been supplemented with a dry dog food containing chondroprotective ingredients (fish oil omega-3 fatty acids, greenlipped mussel meal and glucosamine + chondroitin; Royal Canine Mobility formula) and three dogs had received no supplementation. Three dogs receiving nutritional supplements prior to the test start responded with significant improvement in joint health and mobility and two dogs showed no response suggesting that krill meal alone is in some cases equally or more effective than supplements containing fish oil and/or chondroprotective nutrients (glucosamine + chondroitin).

Example 37

Krill Phospholipid-Peptide Complex Treatment Of Hair Disorders

Twenty individuals are orally administered 1 spoon of either Krill Meal 1 or Krill Meal 2 (PPC) per day for fourteen (14) days. The data is expected to show that PPC administration results in: i) softer hair (50%); and iii) had an overall healthier look (10%). Example 38

Krill Phospholipid-Peptide Complex Treatment Of Nail Disorders Twenty individuals are orally administered 1 spoon of Krill Meal 1 or Krill Meal 2 (PPC) per day for fourteen (14) days. The data is expected to show that PPC administration results in stronger nails (50%).

Example 39

Krill Phospholipid-Peptide Complex Treatment Of Skin Disorders Twenty individuals are orally administered 1 spoon of Krill Meal 1 or Krill Meal 2 (PPC) per day for fourteen (14) days. The data is expected to show that PPC administration result in: i) healthier looking skin (50%); ii) softer skin (50%); and ii) more supple skin (50%).

Example 40

Krill Phospholipid-Peptide Complex Improvement Of Physical Performance

Twenty individuals are orally administered 1 spoon of Krill Meal 1 or Krill Meal 2

(PPC) per day for fourteen (14) days. The data is expected to show that PPC administration results in: i) improved strength (10%); ii) improved stamina (10%); and ii) faster workout recovery (10%). Example 41

Krill Phospholipid-Peptide Complex Treatment Of Skeletomuscular Disorders Twenty individuals are orally administered 1 spoon of Krill Meal 1 or Krill Meal 2 (PPC) per day for fourteen (14) days. The data is expected to show that PPC administration resulted in: i) decreased joint pain (20%); ii) improved joint flexibility (20%). Additionally, one patient is diagnosed with severe joint pain, and undergoing the same PPC administration protocol, reports significant reduction in pain and improved joint function.

Example 42

rill Phospholipid-Peptide Complex Improvement Of Mental Performance

Twenty individuals are orally administered 1 spoon of Krill Meal 1 or Krill Meal 2 (PPC) per day for fourteen (14) days. The data is expected to show that PPC administration resultsin: i) improved concentration (10%); and ii) improved mental acuity (10%). Example 43

Krill Phospholipid-Peptide Complex Treatment Of Multiple Sclerosis Two patients are diagnosed with multiple sclerosis and orally administered 1 spoon of Krill Meal 1 or Krill Meal 2 (PPC) per day for fourteen (14) days. The data is expected to show that, in both patients, PPC administration resulted in: i) dramatic improved strength; ii) dramatically reduced hand numbness.

Example 44

Krill Phospholipid-Peptide Complex Treatment Of Sexual Disorders One patient is diagnosed with impotence and orally administered 1 spoon of Krill Meal 1 or Krill Meal 2 (PPC) per day for fourteen (14) days. The data is expected to show that PPC administration resulted in improved erection and resultant sexual performance.

Claims

Claims We claim:

1. A method, comprising:

a) providing:

i) a patient exhibiting at least one symptom of a medical disorder; ii) a low fluoride crustacean phospholipid-peptide-protein complex (PPC) formulation;

b) administering said formulation to said patient under conditions such that said at least one symptom is reduced.

2. The method of Claim 1, wherein said medical disorder comprises an age-related medical disorder.

3. The method of Claim 2, wherein said age-related medical disorder is selected from at least one of the group consisting of a lack of homeostatic control, macular degeneration, diabetes, or inflammation.

4. The method of Claim 1, wherein said medical disorder comprises malnutrition.

5. The method of Claim 1 , wherein said medical disorder comprises an ocular disorder.

6. The method of Claim 1, wherein said medical disorder comprises a cardiovascular disorder.

7. The method of Claim 1, wherem said medical disorder comprises a skeletal medical disorder.

8. The method of Claim 1, wherein said medical disorder comprises a central nervous system disorder.

9. The method of Claim 1, wherein said medical disorder comprises a muscular disorder.

10. The method of Claim 1, wherein said medical disorder comprises cachexia.

11. The method of Claim 1, wherein said medical disorder comprises digestive tract medical disorder.

12. The method of Claim 1, wherein said medical disorder comprises a dyslipidemic medical disorder.

13. The method of Claim 1 , wherein said medical disorder comprises a hair disorder.

14. The method of Claim 1 , wherein said medical disorder comprises a nail disorder.

15. The method of Claim 1, wherein said medical disorder comprises a skin disorder.

16. The method of Claim 1, wherein said medical disorder comprises a sexual disorder.

17. The method of Claim 1, wherein said formulation is a pharmaceutically acceptable formulation.

18. The method of Claim 1, wherein said PPC formulation is a krill meal formulation.

19. The method of Claim 1 , wherein said PPC formulation is a medicament.

20. The method of Claim 1, wherein said PPC formulation comprises a low fluoride phospholipid-peptide-protein complex.

21. The method of Claim 1, wherein said PPC formulation comprises a low fluoride de- oiled phospholipid-protein complex.

22. The method of Claim 1 , wherein said PPC formulation further comprises a low

fluoride krill oil.

23. The method of Claim 1, wherein said PPC formulation further comprises a low trimethyl amine krill oil.

24. The method of Claim 1, wherein said PPC formulation further comprises a low

fluoride and low trimethyl amine krill oil.

25. The method of Claim 1, wherein said PPC formulation further comprises a low

fluoride extraction residue.

26. The method of Claim 1 , wherein said PPC formulation further comprises an additional ingredient selected from at least one of the group consisting of minerals, lipid soluble vitamins, lipid insoluble vitamins, bioactive health ingredients and omega-3 oils.

27. The method of Claim 1 , wherein said administered composition ranges between 0.005 - 0.50 grams per day per kilogram of said patient's body weight.

28. The method of Claim 1, wherein said formulation is selected from at least one of the group selected from a capsule, a gelatin capsule, a flavored capsule, a tablet, coated tablets, powders, granulates, solutions, dispersions, suspensions, syrups, emulsions, a liquid nutrition composition, a drink and a beverage.

29. The method of Claim 1, wherein said formulation is selected from at least one of the group consisting of a nutritional supplement, a pharmaceutical, a food supplement, a functional food ingredient, a food additive and a nutritional supplement preparation.